In biological research, the cryopreservation of mammalian cells is an invaluable tool. Successful preservation of cells is a top priority given that losing a cell line to contamination or improper storage conditions leads to lost time and money, ultimately delaying research results. Once the cells have been transferred from a cell growth medium to a freezing medium, the cells are typically frozen at a regulated rate and stored in liquid nitrogen vapor or at below -130°C in a mechanical deep freezer. The freeze medium CM-1 enables cryopreservation of cells at below -130°C (or in liquid nitrogen), essentially eliminating the need for an additional, costly ultralow freezer and eliminating time-consuming and demanding controlled rate freezing processes. Simply collect the cells, aspirate the growth medium, resuspend in CM-1, transfer to a cryovial, and store the vial at below -130 °C.
Long shelf-life
CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Trusted by hundreds of researchers
Our advanced cell freezing medium CM-1 is a market-leading product in Germany and Europe and is distinguished by numerous publications involving hundreds of different cell lines worldwide. We tested it with more than 1000 cell lines from our proprietary cell bank.
Optimized ingredients
CM-1 does contain serum products. Serum-containing cryopreservation mediums optimally protect the cells whilst being frozen and have the advantage of high recovery rates. As CM-1 has been tested with a multitude of cell lines, you can rest assured that your cells always recover well.
Contains FBS, DMSO, glucose, salts
Buffering capacity pH = 7.2 to 7.6
Applications & Validation
The cells preserved in our CM-1 freeze medium can be used for cell counting, viability and cryopreservation, cell culture, mammalian cell culture, gene expression analysis and genotyping, in vitro transcription, and polymerase chain reactions. Each batch's efficacy is evaluated using CHO-K1 cells. Each batch is tested for pH, osmolality, sterility, and endotoxins to ensure high quality.
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
What sets DMEM apart from other media is its unique composition. It contains an impressive fourfold increase in amino acid and vitamin concentration compared to the original Eagle's Minimal Essential Medium. Initially developed with low glucose (1 g/L) and sodium pyruvate, DMEM is frequently employed with higher glucose levels, either with or without sodium pyruvate. Notably, DMEM does not contain proteins, lipids, or growth factors, necessitating supplementation. To achieve optimal growth, a common approach is to supplement DMEM with 10% Fetal Bovine Serum (FBS). Additionally, DMEM employs a sodium bicarbonate buffer system (3.7 g/L), requiring a 5-10% CO2 environment to maintain a physiological pH.
Dulbecco's Modified Eagle Medium (DMEM) is highly regarded among defined media for cell and tissue culture, catering to the growth needs of various adherent cell phenotypes. It surpasses the original Eagle's Medium, developed in the 1950s for cultivating chicken cells, through the enhanced supplementary formulation known as Dulbecco's modification. This modification significantly elevates the content of select amino acids and vitamins up to fourfold compared to the original medium.
In the field of cell culture, DMEM plays a vital role as a growth medium for different cell types, including primary cells, stem cells, and transformed cells. Researchers also employ the modified version of DMEM for a wide array of research applications, such as drug discovery, tissue engineering, and the study of cell signaling pathways.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition
Ingredients
Concentration (mg/L)
Inorganic salts
CaCl2 anhydrous
200.00
Fe(NO3)3 ∙ 9H2O
0.10
KCl
400.00
MgSO4 (anhydrous)
97.66
NaCl
6,400.00
NaHCO3
1,500.00
NaH2PO4 ∙ H2O
108.69
Amino acids
L-Arginine ∙ HCl
84.00
L-Cystine ∙ 2HCl
62.58
L-Glutamine
584.00
Glycine
30.00
L-Histidine ∙ HCl ∙ H2O
42.00
L-Isoleucine
104.80
L-Leucine
104.80
L-Lysine ∙ HCl
146.20
L-Methionine
30.00
L-Phenylalanine
66.00
L-Serine
42.00
L-Threonine
95.20
L-Tryptophane
16.00
L-Tyrosine ∙ Na
103.79
L-Valine
93.60
Vitamins
D-Calcium pantothenate
4.00
Choline chloride
4.00
Folic acid
4.00
myo-Inositol
7.00
Nicotinamide
4.00
Pyridoxine ∙ HCl
4.00
Riboflavin
0.40
Thiamine ∙ HCl
4.00
Other components
D (+)-Glucose anhydrous
4,500.00
Sodium pyruvate
110.00
Phenol Red
15.00
This unique formulation combines Dulbecco's Modified Eagle Medium (DMEM) and Ham's F-12 (Ham's Nutrient Mixture F-12) in a precise 1:1 ratio. The addition of L-glutamine further enhances its composition.
DMEM, derived from Eagle's Minimal Essential Medium (EMEM), offers an increased concentration of amino acids and vitamins compared to its predecessor. In contrast, Ham's F-12 is based on Ham's F-10 medium, providing a complementary set of essential components.
To support optimal cell growth, it is common practice to supplement DMEM:Ham's F12 with FBS at a typical concentration of 5-10%. This addition is necessary as the medium lacks growth hormones, lipids, and proteins crucial for cellular development.
DMEM:Ham's F12 incorporates a pH buffer system and is often supplemented with phenol red, a pH indicator. Cultured cells in DMEM:Ham's F12, or any medium utilizing the bicarbonate buffer system, require a controlled CO2 environment of 5-10% to maintain appropriate pH levels. Phenol red enables monitoring of pH changes from 6.2 (yellow) to 8.2 (red).
Quality Control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing as well as warming up to +37°C minimize the quality of the product.
Do not heat the medium to more than 37°C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
This EMEM medium consists of 2 mM L-Glutamine, 1.5 g/L NaHCO3, EBSS, 1 mM Sodium pyruvate, and NEAA.
What's in EMEM?
EMEM is a modified version of Eagle's minimum essential medium, containing Earle's Balanced Salt Solution, non-essential amino acids, L-glutamine, sodium pyruvate, and sodium bicarbonate. It's important to note that this reduced level of sodium bicarbonate (NaHCO3, 1.5 g/L) is intended for use in 5% CO2 in the air. To maintain its effectiveness, storing the medium at two °C to 8°C in the dark when not in use is recommended.
What is EMEM used for?
Eagle's minimal essential medium (EMEM) is a cell culture medium that can maintain cells in tissue culture. The medium contains higher concentrations of amino acids, allowing for a more accurate approximation of the protein composition of cultured mammalian cells. EMEM may be used to cultivate various cells, including fibroblasts, human liver cancer cell line (HepG2) cells and human fetal brain progenitor-derived astrocyte cells (PDA). It is typically used in the presence of fetal bovine serum (FBS), calf, or horse sera.
How is EMEM different from other cell culture media?
While EMEM and Dulbecco's modified Eagle's medium (DMEM) share some similarities, they also differ. Both media lack protein and contain the amino acids, salts, glucose, and vitamins required to provide a cell with energy and maintain it in tissue culture. However, the DMEM formulation is modified to contain up to four times more vitamins and amino acids and two to four times more glucose than EMEM. It's worth noting that EMEM is also different from the original MEM formulation.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
This EMEM medium is formulated with 2 mM L-Glutamine, 2.2 g/L NaHCO3, and EBSS.
What's in EMEM?
EMEM is a modified version of Eagle's minimum essential medium, containing Earle's Balanced Salt Solution, non-essential amino acids, L-glutamine, sodium pyruvate, and sodium bicarbonate. It's important to note that this reduced level of sodium bicarbonate (NaHCO3, 1.5 g/L) is intended for use in 5% CO2 in the air. To maintain its effectiveness, storing the medium at two °C to 8°C in the dark when not in use is recommended.
What is EMEM used for?
Eagle's minimal essential medium (EMEM) is a cell culture medium that can maintain cells in tissue culture. The medium contains higher concentrations of amino acids, allowing for a more accurate approximation of the protein composition of cultured mammalian cells. EMEM may be used to cultivate various cells, including fibroblasts, human liver cancer cell line (HepG2) cells and human fetal brain progenitor-derived astrocyte cells (PDA). It is typically used in the presence of fetal bovine serum (FBS), calf, or horse sera.
How is EMEM different from other cell culture media?
While EMEM and Dulbecco's modified Eagle's medium (DMEM) share some similarities, they also differ. Both media lack protein and contain the amino acids, salts, glucose, and vitamins required to provide a cell with energy and maintain it in tissue culture. However, the DMEM formulation is modified to contain up to four times more vitamins and amino acids and two to four times more glucose than EMEM. It's worth noting that EMEM is also different from the original MEM formulation.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition
Ingredients
Concentration (mg/L)
Inorganic salts
CaCl2 ∙ 2 H2O
264.92
KCl
400.00
Mg SO4
97.67
NaCl
6800.00
NaHCO3
2200.00
NaH2PO4 ∙ H2O
140.00
Amino acids
L-Arginine ∙ HCl
126.00
L-Cystine
24.00
L-Histidine ∙ HCl ∙ H2O
42.00
L-Isoleucine
52.00
L-Leucine
52.00
L-Lysine ∙ HCl
72.50
L-Methionine
15.00
L-Phenylalanine
32.00
L-Threonine
48.00
L-Tryptophane
10.00
L-Tyrosine
36.00
L-Valine
46.00
Vitamins
Choline chloride
1.00
Folic acid
1.00
myo-Inositol
2.00
Thiamine ∙ H2O
1.00
Riboflavin
0.10
D-Calcium Pantothenate
1.00
Nicotinamide
1.00
Pyridoxal ∙ H2O
1.00
Other components
D(+)-Glucose
1000.00
L-Glutamine
292.00
Phenol Red
10.00
Definition of Endothelial Cell Growth Media
An in-vitro cell culture system requires a particular solution called Endothelial Cell Growth Media to sustain the endothelial cells necessary for the experiment. Endothelial cells in the body are exposed to an environment mimicked by the medium, including necessary and non-essential amino acids, vitamins, hormones, growth factors, and trace minerals. This nutrient-dense fluid is critical for preserving the endothelium phenotype and function because it stimulates cell proliferation. Globally, researchers and scientists rely on the CLS Endothelial Cell Growth Medium because it is both effective and simple.
Importance of Endothelial Cell Research
Blood artery creation, blood pressure control, and wound healing are just a few physiological processes in which endothelial cells play a key part. To keep the blood vessels balanced, these cells may react to chemical and mechanical stimuli. In order to fully comprehend the causes of hypertension, atherosclerosis, and thrombosis, research into endothelial cells is crucial. Investigating endothelial cells may also provide light on how to treat these diseases medically. Providing a stable and uniform environment for endothelial cell development and proliferation, the CLS Endothelial Cell Growth Medium has shown to be an indispensable tool in this kind of study.
Quality Assurance
High-quality in-vitro cell culture media is essential for obtaining consistent and repeatable outcomes. Quality control testing is performed on every CLS Endothelial Cell Growth Media batch to guarantee its uniformity and effectiveness. The sterility, lack of mycoplasma and bacteria, and pH levels of each batch are examined. To promote healthy cell development and proliferation, the medium's pH is kept at 7.2 +/
- 0.02, and the temperature is maintained between 20 and 25 degrees Celsius. To ensure the culture is clear of any contamination that might impact the accuracy of the findings, the medium is also checked for microbiological pollutants such as fungus, bacteria, and mycoplasma.
Maintenance and Disclaimer
The expiration date of CLS Endothelial Cell Growth Media is six weeks from the production date. The medium should be kept at a temperature of between +2°C and +8°C, away from light, and never frozen or heated over 37°C to preserve its quality. As a corollary, it is crucial to avoid microwaves and other unregulated heat sources that might compromise the product's integrity. A portion of the medium may be taken out of the container and brought up to room temperature if just a little amount will be utilized. CLS's Endothelial Cell Growth Medium is a tried and true method for cultivating endothelial cells because of the stringent quality controls used in its production.
Notably, the CLS Endothelial Cell Growth Medium is not for clinical or diagnostic use; it is designed primarily for in-vitro usage. Obtaining accurate and reliable findings from the medium requires strict adherence to the manufacturer's guidelines and quality control methods.
Applications of the Medium
The CLS Endothelial Cell Growth Medium has proven useful in several biological studies. As this medium may be used to cultivate endothelial cells, it can be used to simulate endothelial function and malfunction in organ systems in vitro. This is especially useful for studying the blood-brain barrier or tissue-engineered blood arteries. This is essential for understanding disease processes and creating effective treatments.
Endothelial cells, especially HUVEC cells, have been extensively employed in angiogenesis research, wound healing research, and cancer research, thus, the medium has also been modified to promote their proliferation. Accurate and repeatable findings depend on the consistency and high quality of the medium used to develop these cells, and the CLS Endothelial Cell Growth Media meets both of these requirements.
In biological research, the cryopreservation of mammalian cells is an invaluable tool. Successful preservation of cells is a top priority given that losing a cell line to contamination or improper storage conditions leads to lost time and money, ultimately delaying research results. Once the cells have been transferred from a cell growth medium to a freezing medium, the cells are typically frozen at a regulated rate and stored in liquid nitrogen vapor or at below -130°C in a mechanical deep freezer. The freeze medium CM-ACF enables cryopreservation of cells at below -130°C (or in liquid nitrogen), essentially eliminating the need for an additional, costly ultralow freezer and eliminating time-consuming and demanding controlled rate freezing processes. Simply collect the cells, aspirate the growth medium, resuspend in CM-ACF, transfer to a cryovial, and store the vial at below -130 °C.
Long shelf-life
CM-ACF is a serum-free, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Trusted by hundreds of researchers
Our advanced, serum-free cell freezing medium CM-ACF is a market-leading product in Germany and Europe and is distinguished by numerous publications involving hundreds of different cell lines worldwide. We tested it with more than 1000 cell lines from our proprietary cell bank.
Optimized serum-free ingredients
CM-ACF does not contain serum products. Serum-containing cryopreservation mediums have the disadvantage of fluctuating recovery rates and unclear composition. Since the composition and concentration of proteins and other biological components vary from batch to batch in serum, the reproducibility of experiments with cells that were frozen in a serum-containing medium may be compromised. As each component of CM-ACF is carefully defined, you can rest assured that cells always recover identically.
Contains DMSO, glucose, salts
Buffering capacity pH = 7.2 to 7.6
Universal
- even for stem cell preservation
All common cell lines can be frozen and thawed to yield many viable cells. Compared to standard media, the rate of recovery of even the most delicate cells is significantly higher. Using CM-ACF, we store over 1000 different cell lines with outstanding success.
Applications & Validation
The cells preserved in our CM-ACF freeze medium can be used for cell counting, viability and cryopreservation, cell culture, mammalian cell culture, gene expression analysis and genotyping, in vitro transcription, and polymerase chain reactions. Each batch's efficacy is evaluated using CHO-K1 cells. Each batch is tested for pH, osmolality, sterility, and endotoxins to ensure high quality.
One of its notable advantages is the ability to support cell growth without the need for serum supplementation. This eliminates potential interference caused by serum components, ensuring consistent and reliable experimental results. By providing a serum-free culture environment, Ham's F-12 Medium offers researchers greater control over their investigations.
Another key feature of Ham's F-12 Medium is its suitability for single-cell plating. This makes it an excellent choice for a variety of cell lines, including CHO cells, lung cells, and mouse L cells. The medium's optimized nutrient composition facilitates efficient attachment and growth of individual cells, enabling the establishment of homogeneous cell cultures with improved reproducibility.
Moreover, Ham's F-12 Medium has gained recognition as the preferred medium for the Clonal Toxicity Assay (CTA). This assay plays a critical role in assessing the cytotoxic effects of substances on cells. By utilizing Ham's F-12 Medium in the CTA, researchers can accurately evaluate the impact of various compounds or treatments on individual cells, providing valuable insights into toxicological profiles.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Ham's F-12K (Kaighn's) Medium is carefully formulated to optimize cell culture conditions. It features an enriched composition, providing elevated levels of essential components such as amino acids and sodium pyruvate, as well as additional elements including putrescine, thymidine, hypoxanthine, and zinc. These additions enable researchers to supplement the medium with minimal serum or defined components for specific cell types, facilitating precise experimental conditions.
Notably, Ham's F-12K (Kaighn's) Medium does not contain proteins or growth factors. Consequently, supplementation with growth factors and Fetal Bovine Serum (FBS) is often necessary, allowing researchers to tailor the medium to the requirements of their specific cell lines. For optimal performance, the concentration of FBS must be carefully optimized for each cell line, ensuring optimal growth and functionality.
To maintain physiological pH, Ham's F-12K (Kaighn's) Medium employs a sodium bicarbonate buffer system (2.5 g/L), necessitating a controlled 5-10% CO2 environment during cultivation. This ensures the medium's pH remains within the ideal range for cell growth and viability.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing as well as warming up to +37°C minimize the quality of the product.
Do not heat the medium to more than 37°C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium but the basic medium is 8 weeks from the date of manufacture.
IMDM is well suited for rapidly proliferating, high-density cell cultures, including Jurkat, COS-7, and macrophage cells. The various modifications of IMDM available for a range of cell culture applications can be found using the media selector tool. Liquid media provide essential nutrients for all cell culture applications. Each of our high-quality cell culture media is manufactured according to the initially published formula or modifications necessary to the consistent performance and stability of the culture medium.
IMDM vs. DMEM
IMDM contains potassium nitrate instead of ferric nitrate and HEPES and sodium pyruvate. The additional components in IMDM make it more suitable for specialized cell types and specific applications than DMEM.
IMDM vs. RPMI
IMDM and RPMI have different formulations that may be relevant for PMA/ionomycin stimulation. One significant difference is the concentration of Ca2+. While RPMI contains 0.42 mM Ca2+, IMDM contains 1.49 mM.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
One prominent application of McCoy's 5A Medium is its utilization in the culturing of human colon carcinoma cell lines. Specifically, it has been employed in the study of the leucine-rich repeat-containing G-protein-coupled receptor (LGR5) and its role in the metastasis of colon cancer. This medium has been effectively employed in the cultivation of several colon carcinoma cell lines, including HCT116, RKO, FET, CBS, HCT116b, and TENN, enabling researchers to delve deeper into the mechanisms underlying colon cancer metastasis.
In addition to its application in cancer research, McCoy's 5A Medium has proven to be indispensable in the study of osteoblasts. Researchers investigating the ion reactivity of calcium-deficient hydroxyapatite in standard cell culture media have utilized this medium to culture osteoblasts. This application has facilitated a better understanding of the interactions between osteoblasts and calcium-deficient hydroxyapatite, contributing to advancements in the field of bone research.
Notably, McCoy's 5A Medium was meticulously formulated by modifying the amino acids found in Basal Medium Eagle to provide optimal support for liver tumor cells. This enriched formulation enables its suitability for a diverse range of established cell lines, as well as primary cells, further enhancing its versatility and applicability in various research settings.
Moreover, McCoy's 5A Medium extends its biochemical and physiological effects beyond liver tumor cells. It has been successfully employed to support growth in primary cultures of bone marrow, skin, gingiva, kidney, omentum, adrenal, lung, spleen, rat embryo, and other cell types. This wide range of applications attests to the broad utility of McCoy's 5A Medium in supporting the growth and maintenance of various cell types for comprehensive biological research.
Formulation
This McCoy's 5A medium (modified) contains 3.0 g/L of Glucose, stable Glutamine, 2.0 mM of Sodium pyruvate, and 2.2 g/L of NaHCO3.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Medium 199 offers a range of applications in the field. It can effectively maintain the cumulus-oocyte complex (COC) and support the in vitro maturation of oocytes. Additionally, it is employed in the rinsing of aspiration lines during ovum collection from German Holstein cows. Moreover, Medium 199 serves as an excellent medium for the culture of cardiac endothelial cells derived from rats. These applications demonstrate the versatility and adaptability of Medium 199 to various experimental needs.
History
The development of Medium 199 in the 1950s marked a significant advancement in tissue culture media. Prior to its introduction, many culture media relied on animal-derived products and tissue extracts. However, Morgan and colleagues revolutionized the field by formulating a completely defined nutritional source for cell cultures. Through their experiments involving different combinations of vitamins, amino acids, and other factors, they discovered the exceptional growth-promoting properties of Medium 199.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Phosphate-buffered saline (PBS) is a versatile buffer solution used in many biological and chemical applications, as well as tissue processing. Our PBS solution is formulated with high-quality ingredients to ensure a constant pH during experiments. The osmolarity and ion concentrations of our PBS solution are matched to those of the human body, making it isotonic and non-toxic to most cells.
Composition of our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains 137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, and 2 mM KH2PO4. We have chosen this composition based on CSHL protocols and Molecular cloning by Sambrook, which are well-established standards in the research community.
Applications of our PBS Solution
Our PBS solution is ideal for a wide range of applications in biological research. Its isotonic and non-toxic properties make it perfect for substance dilution and cell container rinsing. Our PBS solution with EDTA can also be used to disengage attached and clumped cells. However, it is important to note that divalent metals such as zinc cannot be added to PBS as this may result in precipitation. In such cases, Good's buffers are recommended. Moreover, our PBS solution has been shown to be an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, such as SARS-CoV-2.
Storage of our PBS Solution
Our PBS solution can be stored at room temperature, making it easy to use and access.
To sum up
In summary, our PBS solution is an essential component in many biological and chemical experiments. Its isotonic and non-toxic properties make it suitable for numerous applications, from cell culture to viral transport medium. By choosing our high-quality PBS solution, researchers can optimize their experiments and ensure accurate and reliable results.
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate (2.0 g/L) and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
The unique composition of this RPMI formulation comprises 2.1 mM of stable Glutamine, 2.0 grams per liter of NaHCO3, and phenol red.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate (2.0 g/L) and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
The distinctive formulation of this RPMI includes a concentration of 2.1 mM of stable Glutamine, 2.0 grams per liter of NaHCO3, and is free from phenol red.
Quality Control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate (2.0 g/L) and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
This RPMI 1640 medium contains 4.5 grams per liter of glucose.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Cell culture media: An overview
In the field of life sciences, one of the most important methodologies is cell culture. The removal of cells, tissues, or organs from an animal or plant and the subsequent implantation of those cells, tissues, or organs in an artificial environment that is favorable to their survival and/or growth is what is meant by the phrase "cell culture." Controlled temperature, a substrate for cell attachment, an adequate growth medium, and an incubator that maintains the optimum pH and osmolality are the fundamental environmental needs for optimal cell development. Cells must have these conditions in order to grow to their full potential.
The selection of an adequate growth medium for in vitro cultivation is the stage in cell culture that is both the most critical and the most vital. A growth medium, also known as culture media, is a liquid or gel that has been formulated to encourage the development of organisms on a microscopic, cellular, or plant-like scale. The medium that is used for cultivating cells often contains an adequate supply of energy as well as substances that control the cell cycle. The main components of a culture media include amino acids, vitamins, inorganic salts, glucose, and serum. The serum is added to the medium because it acts as a source of growth factors, hormones, and attachment factors. In addition to providing nutrients, the medium also contributes to the upkeep of pH and osmolality levels.
Types of medium used in cell culture
Both human and animal cells have the potential to be grown in either an artificial or synthetic medium or an entirely natural media that is supplemented with natural elements. In the following, we will give you an overview of the different currently available media types.
Natural media
Only biological fluids that exist in their natural state may be found in natural media. Natural media are very helpful and easy for the cultivation of a broad variety of animal cell types. The lack of understanding of the precise components that make up natural media is the primary factor contributing to the low repeatability of results obtained using natural media.
Artificial media
The preparation of artificial or synthetic media involves the addition of nutrients (both organic and inorganic), serum proteins, carbohydrates, cofactors, vitamins, and salts, as well as O2 and CO2 gas phases [1].
Various types of artificial media have been developed in order to fulfill one or more of the following functions: 1) Immediate survival (a balanced salt solution with a precise pH and osmotic pressure). 2) Prolonged survival (a balanced salt solution supplemented with different formulations of organic chemicals and/or serum). 3) Indefinite development. 4) Specialized functions.
There are four distinct classifications for artificial media:
Serum containing media
The most frequent kind of supplement found in medium used for growing animal cells is fetal bovine serum. It is added to the culture medium as a low-cost supplement in order to achieve the best possible growth conditions. In addition to acting as a transporter or chelator for nutrients that are unstable or water-insoluble, hormones and growth factors, protease inhibitors, and other substances, the serum also binds and neutralizes harmful molecules.
Serum-free Medium
The presence of serum in the media has a number of disadvantages and has the potential to cause major errors in interpretation in immunological research [2, 3]. There have been a variety of different serum-free mediums created [4, 5]. These media are generally specifically formulated to support the culture of a single cell type, such as Knockout Serum Replacement and Knockout DMEM from Thermo Fisher Scientific, and mTESR medium from Stem Cell Technologies [6], for stem cells [7].
Additionally, these media incorporate defined quantities of purified growth factors, lipoproteins, and other proteins, which are otherwise typically provided by the serum [8]. These media are often referred to as "defined cultural media" since the components that make up these media are well understood.
Chemically defined media
These media include ultra-pure inorganic and organic components that have not been contaminated by any kind of contamination. They may also include pure protein additions, such as growth factors.
The genetic modification of bacteria or yeast, together with the addition of particular fatty acids, vitamins, cholesterol, and amino acids, results in the production of their component parts [9].
Protein-free media
Protein-free media are those that do not include any protein at all and instead only include non-protein elements. When compared to media with added serum, the use of media without added protein promotes greater cell proliferation and protein expression and makes it easier to purify any product generated in a downstream process [10-12]. Protein is not included in formulations such as MEM and RPMI-1640. However, a protein supplement might be administered if it is necessary.
Culture media and its basic components
Commercial culture medium may be purchased as a powder or a liquid and often include a variety of nutrients such as amino acids, glucose, salts, vitamins, and other dietary supplements.
The needs for these components are different for each cell line, and these variations are responsible for the wide number of different formulations of media. Each component is responsible for a certain function, which will be outlined in the following paragraphs:
Buffering systems
To maintain optimal growing conditions, pH must be controlled, which is often done by one of two buffering systems:
Natural buffering system
The CO2/H2CO3 ratio in the atmosphere is equal to that of the medium, creating a natural buffering mechanism. In order to preserve their natural buffering mechanism, cultures must be kept in an air environment with 5-10% CO2, which is often achieved by using a CO2 incubator. One of the best things about using a natural buffer is how cheap and safe it is.
HEPES
Chemical buffering using the zwitterion HEPES has a greater buffering capability in the pH range of 7.2-7.4 and does not need a regulated gaseous environment. For particular cell types, a larger dose of HEPES may be harmful. Media containing HEPES are likewise much more susceptible to the phototoxic effects of fluorescent light [13].
Phenol Red
The pH indicator phenol red is often included in commercially available culture medium, allowing for continuous monitoring of pH. By expanding the cells, the metabolites produced by these cells cause a shift in pH and therefore a color change of the media. Phenol red has a dual effect on a medium's color, turning it yellow at acidic pH and purple at alkaline pH. pH 7.4, the optimal value for cell culture, causes the medium to appear fluorescent red.
But phenol red has a few drawbacks: First, phenol red is able to simulate the performance of a number of steroid hormones, primarily estrogen [14]. Therefore, when studying estrogen-sensitive cells like breast tissue, a medium free of phenol red is recommended. Sodium-potassium balance is disrupted by the presence of phenol red in several serum-free formulations. Adding serum or bovine pituitary hormone to the media may counteract this effect [15]. Thirdly, detection in flow cytometric experiments is hindered by the presence of phenol red.
Inorganic salts
Media containing inorganic salts, such as sodium, potassium, and calcium ions, assist maintain osmotic equilibrium and regulate membrane potential.
Amino acids
Since amino acids are the fundamental components of protein, they are an essential component of every single cell growth medium that has ever been conceived. Because cells are unable to produce certain amino acids on their own, it is important that the culture medium include essential amino acids. They are necessary for the proliferation of cells, and the concentration at which they are present determines the maximum cell density that may be attained. In particular, L-glutamine, an essential amino acid is especially crucial.
L-glutamine functions as a secondary source of energy for the metabolism and contributes nitrogen to the production of NAD, NADPH, and nucleotides. Due to the fact that L-glutamine is an unstable amino acid that, with time, changes into a form that cells are unable to utilize, it must be given to the medium.
In addition, non-essential amino acids may be supplied to the medium in order to refuel those that have been used up throughout the growth process. The growth of the cells is boosted and their viability is increased when the growth medium is supplemented with nonessential amino acids.
Carbohydrates
Carbohydrates in the form of sugars are the principal source of energy. Many of the media also include maltose and fructose in addition to the more common sugars of glucose and galactose.
Proteins and peptides
Albumin, transferrin, and fibronectin are the most commonly used proteins and peptides. They are especially significant in media that do not include serum. Albumin, transferrin, aprotinin, fetuin, and fibronectin are some of the proteins that may be found in serum, which is a rich supply of protein.
Albumin is the primary protein found in blood, and its function is to bind and transport various substances, including water, salts, free fatty acids, hormones, and vitamins, between different organs and cells. Albumin's ability to attach to chemicals makes it an effective candidate for removing harmful compounds from the medium in which cells are cultured.
Aprotinin is a protective agent in cell culture systems, since it is stable at neutral and acidic pH, as well as resistant to high temperatures and the destruction that may be caused by proteolytic enzymes. It is capable of inhibiting a number of serine proteases, including trypsin, amongst others.
Fetuin is a glycoprotein that may be detected in higher amounts in the serum of fetal and newborn animals compared to adult serum. In addition to that, it acts as a serine protease inhibitor. The protein fibronectin is an essential component in the process of cell adhesion. Transferrin is a protein that transports iron and is responsible for delivering iron to the membranes of cells.
Fatty acids and lipids
They play a crucial role in serum-free medium when serum is absent.
Vitamins
Numerous vitamins are necessary for cell development and proliferation. Vitamins cannot be produced in adequate amounts by cells and are thus essential in tissue culture as dietary supplements.
In cell culture, the serum is the primary source of vitamins; however, media are also treated with various vitamins to make them suited for a specific cell type. Most typically, the B group vitamins are used for growth stimulation.
Trace elements
Chemical elements such as copper, zinc, selenium, and tricarboxylic acid intermediates are known as trace elements. Trace elements are often added to media that does not include serum in order to replace those that are typically present in serum. These elements are important chemical components that are required for a healthy cell development. Many biochemical reactions depend on certain micronutrients, such as enzyme activity.
Medium supplements
The full growth medium suggested for certain cell lines needs extra components that are absent from the baseline media and serum. These dietary supplements support cell growth and appropriate metabolic function.
Although hormones, growth factors, and signaling molecules are essential for the appropriate proliferation of particular cell lines, the following precautions should always be taken: Since the addition of supplements might alter the osmolality of the complete growth medium, which can inhibit cell development, it is always advisable to verify the osmolality after adding supplements. For the majority of cell lines, the optimum osmolality ranges between 260 and 320 mOSM/kg.
Antibiotics
Antibiotics are often employed to inhibit the development of bacterial and fungal pollutants [16], although they are not essential for cell growth. Since antibiotics might conceal contamination by mycoplasma and resistant bacteria, their routine use is not suggested for cell culture [17, 18].
In addition, antibiotics may disrupt the metabolism of hypersensitive cells. The penicillin-streptomycin combinations made by MilliporeSigma and Life Technologies are often used. Plasmocin has been utilized in the culture of the glioma cell lines TS603, TS516, and BT260 [19], and it has been shown to be effective in removing mycoplasma contamination (20).
Serum
Albumins, growth factors, and growth inhibitors are all present in serum. Serum is one of the most significant components of cell culture medium because it provides amino acids, proteins, vitamins (especially fat-soluble vitamins such as A, D, E, and K), carbohydrates, lipids, hormones, growth factors, minerals, and trace elements.
Serum from fetal and calf bovine sources is often utilized to promote the development of cultured cells. Fetal serum is an abundant supply of growth factors and is suitable for cell cloning and the development of sensitive cells. Due to its diminished growth-promoting capabilities, calf serum is employed in contact-inhibition experiments. Normal growth mediums often include 2% to 10% serum. The addition of serum to culture medium serves the following purposes [21]:
-
The serum delivers the essential nutrients for cells (both in solution and attached to proteins).
-
Several growth factors and hormones involved in growth promotion and specialized cell activity are included in serum.
-
It offers many binding proteins, like albumin and transferrin, that transport other chemicals into the cell. For example, albumin delivers fats, vitamins, hormones, etc. into cells.
-
It also provides proteins, such as fibronectin, which increase cell adhesion to the substrate. Additionally, it produces spreading elements that aid in cell expansion before division.
-
It delivers protease inhibitors that prevent proteolysis in cells.
-
It also contains minerals such as Na+, K+, Zn2+, and Fe2+.
-
It enhances the viscosity of the media, so protecting the cells from mechanical injury during suspension culture agitation.
-
It's also a buffer.
References
[1] Morgan J, Morton H, Parker R. Nutrition of animal cells in tissue culture; initial studies on a synthetic medium. Proc Soc Exp Biol Med. 1950;73:1-8
[2] Kerbel R, Blakeslee D. Rapid adsorption of a foetal calf serum component by mammalian cells in culture. A potential source of artifacts in studies of antisera to cell-specific antigens. Immunology. 1976;31:881-91
[3] Sula K, Draber P, Nouza K. Addition of serum to the medium used for preparation of cell suspensions as a possible source of artifacts in cell-mediated reactions studied by means of the popliteal lymph node test. J Immunogenet. 1980;7:483-9
[4] Mariani E, Mariani A, Monaco M, Lalli E, Vitale M, Facchini A. Commercial serum-free media: hybridoma growth and monoclonal antibody production. J Immunol Methods. 1991;145:175-83
[5] Barnes D, Sato G. Methods for growth of cultured cells in serum-free medium. Anal Biochem. 1980;102:255-70
[6] Yu H, Lu S, Gasior K, Singh D, Vazquez Sanchez S, Tapia O, et al. HSP70 chaperones RNA-free TDP-43 into anisotropic intranuclear liquid spherical shells. Science. 2021;371:
[7] Meharena H, Marco A, Dileep V, Lockshin E, Akatsu G, Mullahoo J, et al. Down-syndrome-induced senescence disrupts the nuclear architecture of neural progenitors. Cell Stem Cell. 2022;29:116-130.e7
[8] Iscove N, Melchers F. Complete replacement of serum by albumin, transferrin, and soybean lipid in cultures of lipopolysaccharide-reactive B lymphocytes. J Exp Med. 1978;147:923-33
[9] Stoll T, Muhlethaler K, von Stockar U, Marison I. Systematic improvement of a chemically-defined protein-free medium for hybridoma growth and monoclonal antibody production. J Biotechnol. 1996;45:111-23
[10] Darfler F. A protein-free medium for the growth of hybridomas and other cells of the immune system. In Vitro Cell Dev Biol. 1990;26:769-78
[11] Barnes D, Sato G. Serum-free cell culture: a unifying approach. Cell. 1980;22:649-55
[12] Hamilton W, Ham R. Clonal growth of chinese hamster cell lines in protein-free media. In Vitro. 1977;13:537-47
[13] Zigler J, Lepe Zuniga J, Vistica B, Gery I. Analysis of the cytotoxic effects of light-exposed HEPES-containing culture medium. In Vitro Cell Dev Biol. 1985;21:282-7
[14] Berthois Y, Katzenellenbogen J, Katzenellenbogen B. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci U S A. 1986;83:2496-500
[15] Karmiol S. Development of serum free media. In: Master JRW, editor. Animal Cell culture, 3rd ed. Oxford:Oxford University Press; 2000.
[16] Perlman D. Use of antibiotics in cell culture media. Methods Enzymol. 1979;58:110-6
[17] McGarrity G. Spread and control of mycoplasmal infection of cell cultures. In Vitro. 1976;12:643-8
[18] Masters J, Stacey G. Changing medium and passaging cell lines. Nat Protoc. 2007;2:2276-84
[19] Chakraborty A, Laukka T, Myllykoski M, Ringel A, Booker M, Tolstorukov M, et al. Histone demethylase KDM6A directly senses oxygen to control chromatin and cell fate. Science. 2019;363:1217-1222
[20] Molla Kazemiha V, Azari S, Amanzadeh A, Bonakdar S, Shojaei Moghadam M, Habibi Anbouhi M, et al. Efficiency of Plasmocin™ on various mammalian cell lines infected by mollicutes in comparison with commonly used antibiotics in cell culture: a local experience. Cytotechnology. 2011;63:609-20
[21] Kragh Hansen U. Molecular aspects of ligand binding to serum albumin. Pharmacol Rev. 1981;33:17-53