Refer to the product datasheet for complete product details.
Briefly, MSCs are cultured in expansion media using the following protocol:
- Plate MSCs in expansion media
- Culture MSCs to 80-90% confluency
- Passage MSCs using fresh expansion media
Reagents supplied in the StemXVivo® Mesenchymal Stem Cell Expansion Media (Catalog # CCM004):
- 250 mL of MSC expansion media
Other Supplies Required
- Penicillin-Streptomycin (100X)
- Trypsin-EDTA (10X)
- Phosphate-Buffered Saline (PBS)
- MSCs (e.g., Rat Mesenchymal Stem Cells, Catalog # PSC003)
- 75 cm2 tissue culture flasks
- 15 mL centrifuge tubes
- Serological pipettes
- Pipettes and pipette tips
- 37 °C and 5% CO2 incubator
- Inverted microscope
- 2 °C to 8 °C refrigerator
- 37 °C water bath
This protocol has been tested using bone marrow- and/or adipose tissue-derived MSCs. If using a different tissue source or cell line, the protocol below may need to be modified.
Culturing Mesenchymal Stem Cells
Add 3.5 - 4.0 x 105 MSCs resuspended in 20 mL of the pre-warmed StemXVivo® MSC Expansion Media to a T75 flask.
Replace the medium every 3 days with fresh StemXVivo MSC Expansion Media.
Culture the MSCs 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.
Add 1-2 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.
Centrifuge at 400 x g for 5 minutes.
Resuspend the cell pellet in 5 mL of StemXVivo® MSC Expansion Media.
Add 3.5-4.0 x 105 MSCs resuspended in 20 mL of pre-warmed StemXVivo® MSC Expansion Media to each T75 flask.