StemXVivo Xeno-Free Human MSC Expansion Media
R&D Systems, part of Bio-Techne | Catalog # CCM021
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, mesenchymal stem cells can be analyzed for marker expression using the following procedure:
- Resuspend cells in pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media
- Plate the cells on Fibronectin-coated plates
- Culture the cells at 37 °C and 5% CO2
Reagents supplied in the StemXVivo® Xeno-Free Human MSC Expansion Media (Catalog # CCM021)::
- 500 mL of Xeno-Free Human MSC expansion media
Reagents and Materials
- Bone marrow-derived MSCs
- Recombinant Human Fibronectin, ACFP (Catalog # ACFP4305)
- Penicillin-Streptomycin (100X)
- TrypLE™ Express (Invitrogen® or equivalent)
- Phosphate-Buffered Saline (PBS)
- 75 cm2 tissue culture flasks
- 15 mL centrifuge tubes
- Serological pipettes
- Pipettes and pipette tips
Equipment
- 37 °C and 5% CO2 humidified incubator
- Centrifuge
- Hemocytometer
- Inverted microscope
- 37 °C water bath
R&D Systems Protocol for Expanding and Subculturing MSCs in StemXVivo® Xeno-Free MSC Expansion Media (Catalog # CCM021)
Culturing Mesenchymal Stem Cells
Coat plates with Recombinant Human Fibronectin.

Pre-warm the required amount of StemXVivo® Xeno-Free Human MSC Expansion Media to room temperature.

Resuspend 4.5-5.0 x 105 cells in 20 mL of the pre-warmed media.
Add the cell suspension to a Fibronectin-coated T75 flask.
Incubate the cells at 37 °C and 5% CO2 in a humidified atmosphere.
Replace spent media every 2-3 days with 20 mL of fresh, pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media.
Subculture the cells when they reach 80-90% confluency. Do not let the cultures become 100% confluent.

Subculturing Mesenchymal Stem Cells
Coat the plates with Recombinant Human Fibronectin.

Pre-warm StemXVivo® Xeno-Free Human MSC Expansion Media and TrypLE Express™ to room temperature.
Remove and discard the media from the flask(s).
Wash the cells once with PBS.
Detach the cells using TrypLE™ Express.

Add 5 mL of StemXVivo® Xeno-Free Human MSC Expansion Media to the flask.
Transfer the cells to a 15 mL conical tube and centrifuge at 400 x g for 5 minutes.
Aspirate off the liquid.
Resuspend the cell pellet in a small amount of pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media.

Perform a cell count.

Resuspend 4.5-5.0 x 105 cells in 20 mL of the pre-warmed StemXVivo® Xeno-Free Human MSC Expansion Media.
Add the cell suspension to a Fibronectin-coated T75 flask.
Incubate the cells at 37 °C and 5% CO2 in a humidified atmosphere.

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