StemXVivo Mouse/Rat Osteogenic Supplement, 12.5 mL
R&D Systems, part of Bio-Techne | Catalog # CCM009
Key Product Details
Summary for StemXVivo Mouse/Rat Osteogenic Supplement, 12.5 mL
Media supplement to induce the differentiation of mouse or rat MSCs into osteocytes. For use with Human/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007).
Key Benefits
- Supports induction of osteogenesis in mouse and rat MSCs
- Defined supplement to reduce experimental variation
- Developed and optimized using mouse and rat MSCs
Why Induce Osteogenesis in MSCs with a Defined Media Supplement?
Despite the well-characterized factors and protocols used to differentiate mesenchymal stem/stromal cells (MSCs) into osteocytes, differentiation efficiencies can vary depending on the quality of the MSC starting population and the reagents used to expand and differentiate MSCs.
StemXVivo® Osteogenic Supplement:
- Contains high quality differentiation factors to drive reproducible and efficient MSC osteogenesis.
- Is defined to reduce unwanted experimental variability.
- Has been developed and optimized using 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
Mouse/Rat StemXVivo® Osteogenic Supplement Components
This media supplement contains high quality factors to drive mouse/rat MSC differentiation into osteocytes.
- This supplement requires media (not included), such as Human/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007) or equivalent.
- The quantity of osteogenic media supplement supplied is sufficient to make 250 mL of media for differentiation.
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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Detection of Osteopontin in Mouse MSC-differentiated Osteocytes. Mouse MSCs were cultured for 14 days using the Human/Mouse/Rat StemXVivo®Osteogenic/Adipogenic Base Media (Catalog # CCM007) and Mouse/Rat StemXVivo®Osteogenic Supplement (Catalog # CCM009). Osteocyte differentiation was verified using a Goat Anti-Mouse Osteopontin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF808). The cells were stained using a NorthernLights™557-conjugated Donkey Anti-Goat Secondary Antibody (Catalog # NL010) and the nuclei were stained with DAPI. |
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Detection of Osteocalcin in Rat MSC-differentiated Osteocytes. Rat MSCs were cultured for 14 days using the Human/Mouse/Rat StemXVivo®Osteogenic/Adipogenic Base Media (Catalog # CCM007) and Mouse/Rat StemXVivo®Osteogenic Supplement (Catalog # CCM009). Osteocyte differentiation was verified using a Mouse Anti-Human Osteocalcin Monoclonal Antibody (Catalog # MAB1419). The cells were stained using a NorthernLights™557-conjugated Donkey Anti-Mouse Secondary Antibody (Catalog # NL010; red) and the nuclei were counterstained with DAPI (blue). |
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, mouse or rat MSCs are differentiated into osteocytes using the following in vitro differentiation procedure:
- Culture multipotent cells of interest
- Induce osteogenic differentiation using a media supplement
- Evaluate differentiation using a mature phenotype marker antibody and fluorescent ICC
For use with Human/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007).
Reagents supplied in the Mouse/Rat StemXVivo® Osteogenic Supplement (Catalog # CCM009):
- 12.5 mL of StemXVivo® Osteogenic Supplement
Note: The quantity of osteogenic media supplement supplied is sufficient to make 250 mL of media for differentiation.
Reagents
- StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007 or equivalent)
- Penicillin-Streptomycin-Glutamate (100X)
Materials
- Mouse or rat MSCs
- 10 cm tissue culture plates
- 15 mL centrifuge tubes
- Pipettes and pipette tips
- Serological pipettes
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 optimized.
Osteogenic Differentiation
Plate 4.2 x 103 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.
Culture cells to 50-70% confluency.
Replace the medium with StemXVivo® Osteogenic Differentiation Medium to induce osteogenesis.
Every 3-4 days, replace with fresh Osteogenic Differentiation Medium.
After 14 days, osteocytes can be harvested and analyzed.
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