Mesenchymal Stem Cell Growth Medium
The Mesenchymal Stem Cell Growth Medium is designed to support the growth of mesenchymal stem/stromal cells from various sources. The medium contains supplements, serum, and antibiotics, eliminating the need for pre-coating the culture vessels with extracellular matrix proteins.
The kit comes with a 450 ml basic growth medium containing phenol red and a 50 ml supplement mix, and the completed medium has a limited shelf life of 4-6 weeks. It is essential to follow the recommended storage conditions, and the kit should not be used for clinical or diagnostic purposes.
Morphology of MSCs propagated in CLS’ Mesenchymal Stem Cell Growth Medium
Figure 1: hDPSC, mesenchymal stem/stromal cells derived from the dental pulp of a donor aged 18 to 25 years cultivated in CLS’ Mesenchymal Stem Cell Growth Medium. Low density on day 1 after seeding (left) and propagated cells on day 3 (right).
Media preparation
- To prepare 500 mL of the complete MSC growth medium, thaw the supplement at room temperature (18-25°C) or overnight at 4-8°C.
- Do not thaw at 37°C in a water bath, which may harm the ingredients.
- Aseptically add the contents of the vials to the Mesenchymal Stem Cell Growth Medium basic.
- We do not recommend using the completed medium for more than 3-4 weeks.
- If other volumes of media are required, prepare accordingly, i.e., 90 ml of Mesenchymal Stem Cell Growth Medium basic plus 10 ml of the supplement mix.
- For sterile filtration purposes, use 0.2 µm low protein binding polyethersulfone (PES) filter units.
Quality control
- pH = 7.2 +/- 0.02 at 20-25°C.
- Each lot has been tested for sterility and the absence of mycoplasma and bacteria.
Maintenance
To ensure proper product maintenance, the basic growth medium should be stored at 4-8°C, and the supplement mix should be stored at -20°C. It is important not to heat the medium to more than 37°C or use uncontrollable heat sources such as microwave appliances, as this may cause damage to the ingredients.
If only a part of the medium is to be used, it should be removed from the bottle and warmed up in a separate device, and the supplement mix should be aliquoted into smaller parts and stored at -20°C. It is also essential to check the expiration dates on the product labels and use the product before the expiration date to ensure its effectiveness.
Multipotency testing of stem cells cultured in CLS’ Mesenchymal Stem Cell Growth Medium
Figure 2: Multipotency of hDPSC as measured by flow cytometry: the cells were propagated in Mesenchymal Stem Cell Growth Medium in regular cell culture flasks and collected at passage 6. Cells were detached using Accutase, rinsed using PBS, and stained for the mesenchymal surface markers CD73, CD90, CD105, and for the absence of the hematopoietic marker HLA-DR. The hDPSCs expressed high levels of the mesenchymal surface markers but lacked the expression of HLA-DR.
Differentiation capability of Mesenchymal stem/stromal cells
Figure 3: Stem cells derived from adipose tissue were cultured in CLS’ Mesenchymal Stem Cell Growth Medium and differentiated using adipogenic differentiation medium (left). For osteogenic differentiation, bone-marrow derived stem cells were cultured in CLS’ Mesenchymal Stem Cell Growth Medium and differentiated using our osteogenic differentiation medium (right).
