Human Mesenchymal Stem Cell Marker Antibody Panel
R&D Systems, part of Bio-Techne | Catalog # SC017
Key Product Details
Summary for Human Mesenchymal Stem Cell Marker Antibody Panel
For the verification of mesenchymal stem/stromal cell identity by flow cytometry.
Key Benefits
- Cost-effective panel of premium quality antibodies
- Defines the starting cell population
- Multiple MSC markers reduce experimental variation
- Efficiently verifies MSC identity
Why is it Important to Verify MSC Identity Using Established Markers?
Researchers use different techniques to isolate, culture, and differentiate human 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 Human Multipotent Mesenchymal Stromal Cell Marker Antibody Panel, which uses expression of multiple established markers to verify MSC identity.
The Human 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 9 markers to increase confidence in MSC identification.
- 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 Human Multipotent Mesenchymal Stromal Cell Marker Antibody Panel includes 9 primary antibodies for the positive and negative identification of Human MSCs by flow cytometry.
Positive MSC Markers
- Mouse Anti-Human Stro-1 IgM gamma Monoclonal Antibody
- Mouse Anti-Human CD44 IgG2A Monoclonal Antibody
- Mouse Anti-Human CD-90 IgG2A Monoclonal Antibody
- Mouse Anti-Human CD105 IgG1 Monoclonal Antibody
- Mouse Anti-Human CD106 IgG1 Monoclonal Antibody
- Mouse Anti-Human CD146 IgG1 Monoclonal Antibody
- Mouse Anti-Human CD166 IgG1 Monoclonal Antibody
Negative MSC Markers
- Mouse Anti-Human CD19 IGg1 Monoclonal Antibody
- Mouse Anti-Human CD45 IGg1 Monoclonal Antibody
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.
Limitations of the Procedure
- FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
- The safety and efficacy of this product in diagnostic or other clinical uses have not been established.
Precautions
- 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.
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Verification of Human Mesenchymal Stem/Stromal Cell Identity by Analysis of MSC Marker Expression. Human mesenchymal stem cells were stained with the primary antibodies included in the Human Multipotent MSC Marker Antibody Panel (Catalog # SC017) (filled histograms) or isotype controls (empty histograms). The cells were then stained with a PE-conjugated Goat Anti-Mouse Secondary Antibody (Catalog # F0102B) and analyzed by flow cytometry. The cells demonstrate the expected phenotype for human MSCs (positive expression of CD44, CD90/Thy1, CD105/Endoglin, CD146/MCAM, CD166/ALCAM, and Stro-1, and negative expression of CD19 and CD45). |
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, human mesenchymal stem cell marker expression can be assessed using the following procedure:
- Incubate cells with the provided antibodies for 30 minutes
- Wash the cells
- Incubate the cells with fluorochrome-conjugated secondary antibodies
- Analyze samples by flow cytometry
Reagents Supplied in the Human Multipotent Mesenchymal Stromal Cell Marker Antibody Panel (Catalog # SC017):
Positive MSC Markers
- Mouse Anti-Human Stro-1 IgM gamma Monoclonal Antibody
- Mouse Anti-Human CD44 IgG2A Monoclonal Antibody
- Mouse Anti-Human CD-90 IgG2A Monoclonal Antibody
- Mouse Anti-Human CD105 IgG1 Monoclonal Antibody
- Mouse Anti-Human CD106 IgG1 Monoclonal Antibody
- Mouse Anti-Human CD146 IgG1 Monoclonal Antibody
- Mouse Anti-Human CD166 IgG1 Monoclonal Antibody
Negative MSC Markers
- Mouse Anti-Human CD19 IGg1 Monoclonal Antibody
- Mouse Anti-Human CD45 IGg1 Monoclonal Antibody
Reagents
- Flow Cytometry Staining Buffer (Catalog # FC001)
- Mouse IgG1 Isotype Control (Catalog # MAB002)
- Mouse IgG2A Isotype Control (Catalog # MAB003)
- Secondary developing reagents (Catalog # F0101B, F0102B, F0103B, F0114, F0116, F0117, F0118, and F0119)
- Sterile PBS
Materials
- Flow Cytometry Tubes
- Serological pipettes
Equipment
- Benchtop centrifuge
Staining of Mesenchymal Stem Cell Surface Markers
Perform a cell count on harvested cells.
Resuspend cells in Flow Cytometry Staining Buffer.
Aliquot 90 μl of the cells into 5 mL flow cytometry tubes.
Add 10 μL of antibody or isotype control (or a previously titrated amount).
Vortex and incubate for 30 minutes at room temperature.
Centrifuge samples at 300 x g for 5 minutes.
Wash the samples three times with Flow Cytometry Staining Buffer.
Resuspend each sample in 200 μL of Flow Cytometry Staining Buffer.
Add 10 μL of a fluorochrome-conjugated secondary antibody (or a previously titrated amount).
Incubate for 30 minutes at room temperature in the dark.
Centrifuge the samples at 300 x g for 5 minutes.
Wash the samples with Flow Cytometry Staining Buffer.
Resuspend the cells in 200-400 μL of Flow Cytometry Staining Buffer.
Analyze the cells by flow cytometry.
