StemXVivo Xeno-Free Human MSC Expansion Media

Catalog # Availability Size / Price Qty
CCM021
Product Details
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StemXVivo Xeno-Free Human MSC Expansion Media Summary

Kit Summary

Xeno-free media optimized for the expansion of human mesenchymal stem cells.

Key Benefits

  • Maintains MSCs in an undifferentiated state
  • Free of animal components that can introduce variability and pose a safety risk
  • Suitable for translational studies
  • Optimized for efficient expansion of human MSCs
 

 

Why use Xeno-Free Media to Expand Mesenchymal Stem Cells?

Mesenchymal stem cells are often expanded in media that contain serum and other components that are of animal origin. The presence of animal-derived factors in media can introduce experimental variability and limit the use of the cultured cells in translational or clinical studies.

R&D Systems offers StemXVivo® Xeno-Free Human MSC Expansion Media, a media that is optimized for efficient expansion of human MSCs in the absence of animal-derived components. MSCs expanded in StemXVivo® Xeno-Free Human MSC Expansion Media maintain a characteristic fibroblast-like morphology, remain in an undifferentiated state, and express MSC markers.

 

 
Mesenchymal Stromal Cells or Mesenchymal Stem Cells?

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® Xeno-Free Human MSC Expansion Media Components

Supplied in a 500 mL volume, StemXVivo Xeno-Free Human MSC Media contains high quality factors to support MSC expansion in vitro. This media does not contain antibiotics.

 

Stability and Storage

StemXVivo® Xeno-Free Human MSC Expansion Media is stable until the expiration date when stored at -20 °C in a manual defrost freezer. The media should be thawed at 2 °C to 8 °C. Thawed media can be aliquoted and stored at -20 °C in a manual defrost freezer for up to 3 months or used within one month when stored in the dark at 2 °C to 8 °C. Avoid repeated freeze-thaw cycles.

 

Data Examples
Morphology of Mesenchymal Stem Cells (MSCs) Cultured in StemXVivo Xeno-Free Human MSC Expansion Media
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Morphology of Mesenchymal Stem Cells (MSCs) Cultured in StemXVivo Xeno-Free Human MSC Expansion Media. Bone marrow-derived human MSCs were cultured in StemXVivo Xeno-Free Human MSC Expansion Media (Catalog # CCM021) for 4 passages and then imaged using brightfield microscopy.
Expression of Mesenchymal Stem Cell (MSC) Markers in MSCs Expanded in R&D Systems StemXVivo Xeno-Free Human MSC Expansion Media or a Competitor's Xeno-Free MSC Expansion Media
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Expression of Mesenchymal Stem Cell (MSC) Markers in MSCs Expanded in R&D Systems StemXVivo® Xeno-Free Human MSC Expansion Media or a Competitor’s Xeno-Free MSC Expansion Media. Human bone marrow-derived MSCs were expanded for 4 passages in StemXVivo® Xeno-Free Human MSC Expansion Media (Catalog # CCM021) or a competitor’s xeno-free MSC expansion media. The cells were harvested and stained for the expression of positive and negative MSC markers. Positive MSC markers were detected with APC-conjugated Mouse Anti-Human Endoglin/CD105 Monoclonal Antibody (Catalog # FAB10971A), PE-conjugated Mouse Anti-Human 5’-Nucleotidase/CD73 Monoclonal Antibody (Catalog # FAB5795P), and APC-conjugated Mouse Anti-Human CD90/Thy1 Monoclonal Antibody (Catalog # FAB2067A). Expression of the negative MSC marker, CD45 was detected with PE-conjugated Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # FAB1430P). Quadrants were set based on isotype controls.
 

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.

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Mesenchymal Stem Cells

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.

Alternate Names
AEG1; LYRIC; LYRIC/3D3; Mesenchymal Stem Cells; metadherin

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 Provided

Reagents supplied in the StemXVivo® Xeno-Free Human MSC Expansion Media (Catalog # CCM021)::

  • 500 mL of Xeno-Free Human MSC expansion media

 

Other Supplies Required

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

 

Protocol Overview

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.

Coat plates with Recombinant Human Fibronectin.

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

Pre-warm the required amount of media.

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.

Resuspend cells in 20 mL of the pre-warmed media.
 

Subculturing Mesenchymal Stem Cells

Coat the plates with Recombinant Human Fibronectin.

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.

Pre-warm media.

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.

Add 5 mL media to the flask.

Perform a cell count.

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.

Resuspend cells in pre-warmed media.

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