StemXVivo Mesenchymal Stem Cell Expansion Media Best Seller
R&D Systems, part of Bio-Techne | Catalog # CCM004
Key Product Details
Summary for StemXVivo Mesenchymal Stem Cell Expansion Media
A complete, fetal bovine serum-containing media formulated and optimized for the maintenance and expansion of purified human, mouse, and rat MSCs.
Key Benefits
- Formulated to reduce variation
- All components, including fetal bovine serum, were selected and optimized for MSCs
- Ready to use
Why Culture MSCs in a Defined Medium?
Mesenchymal stem/stromal cells (MSCs) are a rare population of multipotent cells that can be derived from a number of tissues such as bone marrow, adipose, and placenta.
Given their rarity and sensitivity to ex vivo conditions, MSCs are most often expanded in vitro under specific culture conditions.
StemXVivo® MSC Expansion Media:
- Supports reproducible MSC growth in vitro.
- Can be supplemented with user-defined cytokines and growth factors, although these are not needed for culture
- Has been developed and optimized for use with human, mouse, and rat MSCs.
The term ‘mesenchymal stromal cells’ is commonly used to describe a heterogeneous population of cultured cells that are adherent to plastic, have a distinct morphology, and express a specific set of marker proteins. Within this heterogeneous population are cells referred to as ‘mesenchymal stem cells.’
Mesenchymal stem cells are multipotent, self-renewing cells that have the ability to differentiate into adipocytes, chondrocytes, and osteoblasts when cultured in vitro. Read More about MSC Nomenclature
StemXVivo® MSC Expansion Media Components
Supplied in a 250 mL volume, this medium contains high quality factors to support MSC expansion in vitro.
- Supplemented with sodium bicarbonate
- Does not contain antibiotics
2006 Proposed Change to MSC Nomenclature
Although mesenchymal stromal cells were once referred to as ‘mesenchymal stem cells’, a change to ‘mesenchymal stromal cells’ was proposed by the International Society for Cellular Therapy in 2006.1
The change in nomenclature originates from two important factors:
- Methods used to isolate mesenchymal stem cells yield a heterogeneous population of cells with only a fraction of these cells demonstrating multipotency.
- The absence of direct evidence that mesenchymal stem cells can self-renew and differentiate in vivo.
Use of Mesenchymal Stem and Stromal Cell Terminology
Data supporting MSC self-renewal and multipotency have been obtained using in vitro conditions, which does not adequately reflect the in vivo environment. The lack of in vivo data has led some researchers to question the validity of the term ‘mesenchymal stem cell’ providing further support for the use of ‘mesenchymal stromal cells’ to describe MSCs.2 While ‘mesenchymal stromal cells’ may be the more scientifically accurate term for MSCs, the two terms are often used interchangeably in the literature. R&D Systems recognizes the use of both mesenchymal stem cells and mesenchymal stromal cells and uses ‘MSC’ to indicate mesenchymal stem/stromal cells to account for both designations.
Definitions of Mesenchymal Stromal Cells and Mesenchymal Stem Cells
- Mesenchymal Stromal Cells – A heterogeneous population of cultured cells with similar characteristics such as the ability to adhere to plastic and the expression of specific marker proteins.
- Mesenchymal Stem Cells – A subpopulation of mesenchymal stromal cells that have the capacity to self-renew and differentiate into mesodermal lineages when cultured in vitro. The capacity to self-renew and differentiate in vivo has yet to be clearly demonstrated for mesenchymal stem cells.
References
- Dominici, M. et al. (2006) Cytotherapy 8:315.
- Keating, A. (2012) Cell Stem Cell 10:709.
The term 'mesenchymal stem cells' (MSCs) is most commonly used to describe multipotent self-renewing cells that can be differentiated in vitro to generate adipocytes, chondrocytes, and osteoblasts. However, because these biological properties and hierarchical relationships remain to be clearly demonstrated in vivo, the term 'multipotent mesenchymal stromal cells' is often used to distinguish cultured cells from their in vivo precursors. Originally discovered in mouse bone marrow, multipotent mesenchymal stromal cells cultured from a variety of species and tissue types, have been shown to differentiate into progeny of additional lineages including, cardiomyocytes, endothelial cells, hepatocytes, and neural cells. Again, the physiological relevance of these findings remains to be determined.
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Phenotypic Analysis of Human MSCs Expanded in StemXVivo® MSC Expansion Media. Human MSCs were expanded using StemXVivo®Mesenchymal Stem Cell (MSC) Expansion Media (Catalog # CCM004). Filled histograms indicate cells stained with markers of undifferentiated MSCs including anti-CD105 (Catalog # FAB10971P) or CD45 (Catalog # MAB1430). The open histograms show isotype-matched control staining. MSCs appropriately lack expression of CD45. |
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Phenotypic Analysis of Mouse MSCs Expanded in StemXVivo® MSC Expansion Media. Mouse MSCs were expanded using StemXVivo®Mesenchymal Stem Cell (MSC) Expansion Media (Catalog # CCM004). Filled histograms indicate cells stained with markers of undifferentiated MSCs including anti-CD44 (Catalog # AF6127) or CD45 (Catalog # MAB114). The open histograms show isotype-matched control staining. MSCs appropriately lack expression of CD45. |
Assay Procedure
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
Reagents
- Penicillin-Streptomycin (100X)
- Trypsin-EDTA (10X)
- Phosphate-Buffered Saline (PBS)
Materials
- MSCs (e.g., Rat Mesenchymal Stem Cells, Catalog # PSC003)
- 75 cm2 tissue culture flasks
- 15 mL centrifuge tubes
- Serological pipettes
- Pipettes and pipette tips
Equipment
- 37 °C and 5% CO2 incubator
- Centrifuge
- Hemocytometer
- 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.
Perform a cell count.
Add 3.5-4.0 x 105 MSCs resuspended in 20 mL of pre-warmed StemXVivo® MSC Expansion Media to each T75 flask.