StemXVivo Serum-Free Human MSC Expansion Media
R&D Systems, part of Bio-Techne | Catalog # CCM014
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, MSCs are cultured in serum-free expansion media using the following protocol:
- Plate MSCs on Fibronectin-coated culture dishes
- Culture MSCs to 80-90% confluency in serum-free expansion media
- Passage MSCs using fresh serum-free expansion media
Reagents supplied in the StemXVivo® Serum-Free Mesenchymal Stem Cell Expansion Media (Catalog # CCM014):
- 250 mL of serum-free MSC expansion media
Reagents
- Penicillin-Streptomycin (100X)
- Human Fibronectin (R&D Systems, Catalog # 1918-FN-02M)
- Trypsin-EDTA (10X)
- Phosphate-Buffered Saline (PBS)
Materials
- Human MSCs
- 75 cm2 tissue culture flasks
- 15 mL centrifuge tubes
- Pipettes and pipette tips
- Serological pipettes
Equipment
- 37 °C and 5% CO2 incubator
- Centrifuge
- Hemocytometer
- Inverted microscope
- 2 °C to 8 °C refrigerator
- 37 °C water bath
Culturing Mesenchymal Stem Cells
Coat plates with 5 µg/mL Human Fibronectin.
Add 4.5-5.0 x 105 MSCs resuspended in 20 mL of the pre-warmed StemXVivo® Serum-Free MSC Expansion Media to a T75 flask.
Replace the medium every 2-3 days with fresh StemXVivo® Serum-Free MSC Expansion Media.
Culture cells to 80-90% confluency.
Subculturing Mesenchymal Stem Cells
Remove and discard the media from the T75 tissue culture flask(s) containing the MSCs.
Wash the cells with PBS.
Thawing of Cryopreserved Stem Cells
Add 3 mL of pre-warmed trypsin to the MSCs and incubate the flask at 37 °C.
Transfer the dissociated MSCs to a 15 mL conical tube and centrifuge at 400 x g for 5 minutes.
Resuspend the cell pellet in 5 mL of StemXVivo® Serum-Free MSC Expansion Media.
Perform a cell count..
Add 4.5-5.0 x 105 MSCs resuspended in 20 mL of pre-warmed StemXVivo® Serum-Free MSC Expansion Media into each Fibronectin-coated T75 flask.
Culture the MSCs to 80-90% confluency.
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