21/07/2023

INS-1

INS-1 is a well-characterized rat insulinoma cell line widely used in diabetes research. INS-1 cells secrete insulin upon glucose stimulation; therefore, they are employed to study glucose metabolism, beta cell physiology, and insulin secretion regulation. Further, these cells are also utilized in screening, testing, and developing potential therapeutics against diabetes mellitus.

This article will provide you with all the necessary information about INS-1 beta cells you need to know before using them in your research. Mainly, you will learn the following:

  1. General characteristics and origin of INS-1 cells
  2. INS-1 cell line: Culturing information
  3. Advantages & disadvantages of INS-1 cell line
  4. Research applications of INS-1 rat insulinoma cells
  5. Research publications featuring INS-1 beta cells
  6. Resources for INS-1 Cell line: Protocols, Videos, and More

 1.      General characteristics and origin of INS-1 cells

Knowing about a cell line's general features and origin can greatly help you efficiently and effectively use it in your research. This article section will inform you about INS-1 origin and general characteristics. You will know: What is the INS-1 rat Insulinoma cell line? What are the general characteristics of INS-1? What is INS-1 832/3 cell line? What is INS-1E?

  • INS-1 cells were originally isolated from a 666-day-old rat with X-ray-induced transplantable insulinoma.
  • INS-1 cells are bi-hormonal. They simultaneously express insulin and proglucagon proteins. These cells are considered immature as they exhibit low expression levels of the Nkx6.1 transcription factor and are deprived of alpha cell markers [1].
  • There are two subclones of INS-1 cells, i.e., INS-1E and INS-1 832/3.
  • INS-1E differs from parent cell line INS-1 regarding secretary responses to glucose and insulin content.
  • INS-1 832/3 or INS-1 832/13 is also a subclone of the INS-1 cell line. It is an invaluable model for studying the function of pancreatic islet beta cells and insulin secretion regulation. It also differs from parent INS-1 cells in the context of glucose-stimulated insulin secretions (GSIS).

3D modeling of insulin-mediated glucose uptake.

2.      INS-1 cell line: Culturing information

To efficiently handle and maintain a cell line, you must know the following culturing information about it. This section of the article will cover all the key points for culturing INS-1 cells. you will learn: How do you culture INS-1 beta cells? What is INS-1 cell culture protocol? What is the doubling time of INS-1 cells? What is the INS-1 rat insulinoma cell media?

Key Points for Culturing INS-1 Cells

Doubling Time:

The population doubling time of INS-1 cells is approximately 44 hours.

Adherent or in Suspension:

INS-1 cells grow both in suspension and adherent form.

Sub-cultivation Ratio:

INS-1 cells are sub-cultured at a split ratio of 1:3. Briefly, suspended cells are collected. Adherent cells are rinsed with PBS and incubated with the Accutase solution. After detachment cells are added with fresh media. Afterwards, both suspended and adherent cells are centrifuged and collected. Cells are carefully resuspended and dispensed into the new flasks for growth.

Growth Medium:

RPM1 1640 is used to culture the INS-1rat insulinoma cell line. The media is supplemented with 10% heat-inactivated fetal bovine serum, 2.1 mM stable Glutamine, 10 mM HEPES, 2.0 g/L NaHCO3, and 1 mM sodium pyruvate.

Growth Conditions:

INS-1 cells are kept in a humidified incubator set at 37°C temperature and with a continuous 5% CO2 supply.

Storage:

INS-1 beta cells can be stored in the vapour phase of liquid nitrogen or at below -150°C temperature in an electric freezer for the long term.

Freezing Process and Medium:

CM-1 or CM-ACF media are used to freeze INS-1 cells through a slow freezing process. It allows only a 1°C drop in temperature per minute to protect cell viability.

Thawing Process:

Frozen INS-1 cells are thawed in a water bath pre-set at 37 degrees Celsius temperature for 40 to 60 seconds. After thawing, cells are added with fresh media and directly poured into a new flask for growth. After 24 hours media is replaced to eliminate freezing media components.

Biosafety Level:

Biosafety level 1 laboratory is required to culture INS-1 rat insulinoma cells.

 

INS-1 cells growing in islets at 10x and 20x magnification.

3.      Advantages & disadvantages of INS-1 cell line

Like other cell lines, INS-1 also possesses some distinct features associated with certain pros and cons. Herein, we have mentioned a few significant ones.

Advantages

The main advantages of the INS-1 cell line are:

Well-characterized

INS-1 is a well-established and well-characterized cell line. It has been used in numerous research studies. It maintains its phenotypic characteristics and insulin secretion capacities over a longer period, thus providing reliable and consistent experimental outcomes.

Beta cell model

INS-1 cells are used to study pancreatic islet beta cells' function as they secrete insulin and are responsive to fluctuations in glucose levels.

 

Disadvantages

The disadvantages of INS-1 cells are:

Non-human origin

INS-1 beta cells have a non-human origin. They were derived from rat insulinoma. This may cause species-specific differences and limit the direct translation of experimental results to human physiology.

 

4.      Research applications of INS-1 rat insulinoma cells

The INS-1 beta cells are extensively used in diabetes research. A few promising applications of this cell line are mentioned here.

  • Insulin secretion studies: INS-1 cells have insulin-secreting ability and thus are extensively employed to study the underlying cellular mechanisms of insulin secretion. Researchers investigate essential factors involved in insulin release, including glucose metabolism, signalling pathways, hormones, and pharmacological agents. A study found that a K + ATP ion channel-dependent pathway regulates insulin secretion in INS-1 beta cells [2]. Besides, studies have also revealed that GLP-1R and AKT/PDX1 pathways are also involved in insulin secretion in INS-1 rat insulinoma cells [3].
  • Beta cell function studies: INS-1 cells have pancreatic islet beta cell-like characteristics such as responsiveness to glucose metabolism and insulin secretion. Therefore, they are used to study beta cells' physiological processes and functions. A study conducted in 2022 utilized INS-1 cells and developed a beta cell dysfunction model through H2O2 They studied the cell viability, insulin secretion, and oxidative stress-related markers in these cells in response to natural compound treatment [4].
  • Drug discovery and development: The rat insulinoma cells INS-1 are widely used to screen and test anti-diabetic compounds or drugs. They can be employed to study the potential effects of therapeutic agents on insulin secretion and other relevant parameters. A study found that loganin, a component of a Chinese herbal formula, protected the insulin-secreting function of INS-1 cells and exerted potential anti-diabetic effects. The component mediated these beneficial effects by inhibiting the nuclear translocation of the FOXO1 gene through the PI3K/AKT pathway [5].

5.      Research publications featuring INS-1 beta cells

Here are some significant research publications featuring the INS-1 rat insulinoma cell line.

Alpha-mangostin improves insulin secretion and protects INS-1 cells from streptozotocin-induced damage

This study was published in the Internation Journal of Molecular Sciences in 2018. The study proposed that a yellow crystalline natural compound, Alpha-mangostin, promotes insulin secretion in INS-1 beta cells and protects them against damage induced by beta cell toxin streptozotocin.

Epicatechin potentiation of glucose-stimulated insulin secretion in INS-1 cells is not dependent on its antioxidant activity

This research was published in acta pharmacologica sinica in 2018. The study finding revealed that the epicatechin compound promotes glucose-stimulated insulin secretion in saturated fatty acid-impaired INS-1 beta cells via CaMKII pathway activation.

A Newly Discovered Phenylethanoid Glycoside from Stevia rebaudiana Bertoni Affects Insulin Secretion in Rat INS-1 Islet β Cells

This article in the Molecules (2019) proposed that a novel natural compound Phenylethanoid Glycoside affects insulin secretion in INS-1 rat insulinoma cells, and thus has anti-diabetic potential.

Loureirin B promotes insulin secretion through GLP-1R and AKT/PDX1 pathways

This research was published in the European Journal of Pharmacology (2022). The study proposed that Loureirin B, a natural product enhances insulin secretion in INS-1 pancreatic islet beta cells via modulation of  AKT/PDX1 and GLP-1R pathways.

In vitro anticancer activities of Withania coagulans against HeLa, MCF-7, RD, RG2, and INS-1 cancer cells and phytochemical analysis

This Integrative Medicine Research (2018) article evaluated the anticancer potential of Withania coagulans extract using INS-1 cancer cells.

6.      Resources for INS-1 Cell line: Protocols, Videos, and More

Here are some online resources featuring the INS-1 cells:

The following link contains the INS-1 cell culture protocol:

  • INS-1 cell line: This website contains all the information about INS-1 cell culture. It includes INS-1 cell culture and freezing media information, protocols for subculturing, and handling of cryopreserved and proliferative INS-1 cultures.

References

  1. Acosta-Montalvo, A., et al., Proglucagon-Derived Peptides Expression and Secretion in Rat Insulinoma INS-1 Cells. Front Cell Dev Biol, 2020. 8: p. 590763.
  2. Park, J.E. and J.S. Han, A Portulaca oleracea L. extract promotes insulin secretion via a K+ ATP channel dependent pathway in INS-1 pancreatic β-cells. Nutrition Research and Practice, 2018. 12(3): p. 183.
  3. Fang, H., et al., Loureirin B promotes insulin secretion through GLP-1R and AKT/PDX1 pathways. European Journal of Pharmacology, 2022. 936: p. 175377.
  4. Duan, J., et al., Swietenine and swietenolide from Swietenia macrophylla king improve insulin secretion and attenuate apoptosis in H2O2 induced INS‐1 cells. Environmental Toxicology, 2022. 37(11): p. 2780-2792.
  5. Mo, F.-F., et al., Anti-diabetic effect of loganin by inhibiting FOXO1 nuclear translocation via PI3K/Akt signaling pathway in INS-1 cell. Iranian Journal of Basic Medical Sciences, 2019. 22(3): p. 262.