Mouse Mesenchymal Stem Cell Marker Antibody Panel

For the verification of mouse mesenchymal stem/stromal cell identity by flow cytometry.

Why is it Important to Verify MSC Identity Using Established Markers?

Researchers use different techniques to isolate, culture, and differentiate mesenchymal stem/stromal cells (MSCs). Variations in experimental approaches as well as differences in the MSC starting population may account for experimental variability and some of the contradictory data that have been published in this field.

One way to minimize experimental variation is to clearly define the starting cell population by using a functional or phenotypic assay. R&D Systems offers the Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel, which uses expression of multiple established markers to assess MSC identity.

The Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel:

• Enables verification of MSC identity in less than 2 hours (with use of fluorochrome-conjugated secondary antibody).
• Includes antibodies for the positive and negative identification of MSCs.
• Uses 8 markers to increase confidence in MSC identification by flow cytometry.
• Provides a faster method to verify MSC identity compared to differentiation protocols.

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

The Mouse Multipotent Mesenchymal Stromal Cell Marker Antibody Panel includes 8 primary antibodies for the positive and negative identification of Mouse MSCs by flow cytometry.

Positive MSC Markers

• Rat Anti-Mouse Sca-1 IgG_2A Monoclonal Antibody
• Rat Anti-Mouse CD29 IgG_2A Monoclonal Antibody
• Rat Anti-Mouse CD44 IgG_2B Monoclonal Antibody
• Rat Anti-Mouse CD73 IgG_2A Monoclonal Antibody
• Rat Anti-Mouse CD105 IgG_2A Monoclonal Antibody
• Rat Anti-Mouse CD106 IgG_2A Monoclonal Antibody

Negative MSC Markers

• Rat Anti-Mouse CD11b IgG_2B Monoclonal Antibody
• Rat Anti-Mouse CD45 IgG_2B Monoclonal Antibody

Each antibody is supplied as a 50X stock; enough for 25 assays when used in 500 µL staining volume per assay. Each 1X antibody solution has an endotoxin level of

Stability and Storage

Store the unopened kit at 2 °C to 8 °C. Use within 1 year of receipt.

Opened/Reconstituted reagents may be stored for up to 1 month at 2 °C to 8 °C.
* Aliquot and store at =-20 °C in a manual defrost freezer for up to 6 months.
* Avoid repeated free-thaw cycles.
* Provided this is within 1 year from receipt.

PRECAUTION

• The acute and chronic effects of over-exposure to reagents in this kit are unknown.
• Safe laboratory procedures should be followed and protective clothing should be worn when handling kit reagents.

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.