StemXVivo Xeno-Free Human MSC Expansion Media
Catalog # CCM021 | R&D Systems, Inc. a Bio-Techne Brand
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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FAQs for StemXVivo Xeno-Free Human MSC Expansion Media
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Is the appearance of crystals in StemXVivo® Xeno-free Human MSC Expansion Media (Catalog # CCM021) normal?
Yes. The crystals are a calcium-containing precipitate and may form, to a limited degree, in this media. A limited amount of crystals will not impact MSC expansion. Avoiding multiple freeze-thaw cycles of the medium is helpful in reducing the amount of crystals.
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Does StemXVivo Xeno-Free Human MSC Expansion Media, catalog # CCM021 contain purified Albumin?
Yes, there is purified Human Serum Albumin (HSA) in the media. We have found it is essential for optimal performance and have not identified an alternative. We do not offer an albumin-free MSC media. CCM021 is Serum free and Xeno-free, but still contains human-derived components.