StemXVivo Human Osteogenic Supplement, 12.5 mL

R&D Systems | Catalog # CCM008

R&D Systems
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Key Product Details

Species

Human

Product Summary for StemXVivo Human Osteogenic Supplement, 12.5 mL

Kit Summary

Media supplement to induce the differentiation of human MSCs into osteocytes. For use with Human/Mouse/Rat StemXVivo Osteogenic/Adipogenic Base Media (Catalog # CCM007).

Key Benefits

  • Supports induction of osteogenesis in human MSCs
  • Defined supplement to reduce experimental variation
  • Developed and optimized using human 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.
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

Human StemXVivo® Osteogenic Supplement Components

This media supplement contains high quality factors to drive human 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.
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Scientific Data Images for StemXVivo Human Osteogenic Supplement, 12.5 mL

Detection of Osteocalcin in Human MSC-differentiated Osteocytes.

Detection of Osteocalcin in Human MSC-differentiated Osteocytes.

Human MSCs were differentiatedin vitrofor 14 days using Human/Mouse/Rat StemXVivo®Osteogenic/Adipogenic Base Media (Catalog # CCM007) and Human StemXVivo®Osteogenic Supplement (Catalog # CCM008). Osteocyte differentiation was verified using a Mouse Anti-Human Osteocalcin Monoclonal Antibody (Catalog # MAB1419). The cells were stained with a NorthernLights(NL)557-conjugated Donkey Anti-Mouse Secondary Antibody (Catalog # NL007; red), and the nuclei were counterstained with DAPI (blue).

Formulation, Preparation, and Storage

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

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, metadherin

Additional Mesenchymal Stem Cells Products

Product Documents for StemXVivo Human Osteogenic Supplement, 12.5 mL

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for StemXVivo Human Osteogenic Supplement, 12.5 mL

For research use only

Citations for StemXVivo Human Osteogenic Supplement, 12.5 mL

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Protocols

View specific protocols for StemXVivo Human Osteogenic Supplement, 12.5 mL (CCM008):

Refer to the product datasheet for complete product details.

Briefly, human 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).
View Graphic Procedure

 

 

Reagents Provided

plied in the Human/

Reagents supplied in the Human StemXVivo® Osteogenic Supplement (Catalog # CCM008):

  • 12.5 mL of StemXVivo® Osteogenic Supplement

    Note: The quantity of osteogenic media supplement is sufficient to make 250 mL of media for differentiation.

 

Other Supplies Required

Reagents

  • StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007 or equivalent)
  • Penicillin-Streptomycin-Glutamate (100X)

Materials

  • Human 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

 

Procedure Overview

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

  1. Plate 4.2 x 103 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.
  2. Culture cells to 50-70% confluency.
  3. Replace the medium with StemXVivo® Osteogenic Differentiation Medium to induce osteogenesis.
  4. Every 3-4 days, replace with fresh Osteogenic Differentiation Medium.
  5. After 14 days, osteocytes can be harvested and analyzed.

FAQs for StemXVivo Human Osteogenic Supplement, 12.5 mL

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  • Q: In the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006), are Part #'s 90415, 390416, and 390417 the same as the StemXVivo® Human Adipogenic Supplement (Catalog # CCM011), StemXVivo® Human Osteogenic Supplement (Catalog # CCM008), and StemXVivo® Human Chondrogenic Supplement (Catalog # CCM006), respectively?

    A: Yes, the StemXVivo® Human Adipogenic Supplement (Catalog # CCM011), StemXVivo®  Human Osteogenic Supplement (Catalog # CCM008), and StemXVivo® Human Chondrogenic Supplement (Catalog # CCM006) are the same as Part #'s 390415, 390416, and 390417, respectively, in the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006).

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