StemXVivo Human Osteogenic Supplement, 12.5 mL

Catalog # Availability Size / Price Qty
CCM008
Product Details
Procedure
Citations (11)
FAQs
Supplemental Products
Reviews (1)

StemXVivo Human Osteogenic Supplement, 12.5 mL Summary

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.

 

Data Examples
Detection of Osteocalcin in Human MSC-differentiated Osteocytes
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Detection of Osteocalcin in Human MSC-differentiated Osteocytes. Human MSCs were differentiated in vitro for 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).

 

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.

Specifications

Shipping Conditions
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.
Species
Human

Product Datasheets

Assay Procedure

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).

 

 

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

Plate 4.2 x 103 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.

Culture cells to 50-70% confluency.

Protocol for Human CD4+ T Cell Enrichment Column

Replace the medium with StemXVivo® Osteogenic Differentiation Medium to induce osteogenesis.

Protocol for Human CD4+ T Cell Enrichment Column

Every 3-4 days, replace with fresh Osteogenic Differentiation Medium.

 

After 14 days, osteocytes can be harvested and analyzed.

Protocol for Human CD4+ T Cell Enrichment Column

Reagents Provided

Citations for StemXVivo Human Osteogenic Supplement, 12.5 mL

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

11 Citations: Showing 1 - 10
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  1. BMP4 and perivascular cells promote hematopoietic differentiation of human pluripotent stem cells in a differentiation stage-specific manner
    Authors: S Jeong, B An, JH Kim, HW Han, JH Kim, HR Heo, KS Ha, ET Han, WS Park, SH Hong
    Exp. Mol. Med., 2020;0(0):.  2020
  2. Regulation of Mesenchymal Stem Cell Differentiation by Nanopatterning of Bulk Metallic Glass
    Authors: AM Loye, ER Kinser, S Bensouda, M Shayan, R Davis, R Wang, Z Chen, UD Schwarz, J Schroers, TR Kyriakides
    Sci Rep, 2018;8(1):8758.  2018
  3. An immortalised mesenchymal stem cell line maintains mechano-responsive behaviour and can be used as a reporter of substrate stiffness
    Authors: A Galarza To, JE Shaw, A Wood, HTJ Gilbert, O Dobre, P Genever, K Brennan, SM Richardson, J Swift
    Sci Rep, 2018;8(1):8981.  2018
  4. Differentiation of human iPSCs into VSMCs and generation of VSMC-derived calcifying vascular cells
    Authors: A Trillhaase, U Haferkamp, A Rangnau, M Märtens, B Schmidt, M Trilck, P Seibler, R Aherrahrou, J Erdmann, Z Aherrahrou
    Stem Cell Res, 2018;31(0):62-70.  2018
  5. Development of a Three-Dimensional Bioengineering Technology to Generate Lung Tissue for Personalized Disease Modeling
    Stem Cells Transl Med, 2016;0(0):.  2016
  6. Human fetal and adult bone marrow-derived mesenchymal stem cells use different signaling pathways for the initiation of chondrogenesis.
    Authors: Brady K, Dickinson S, Guillot P, Polak J, Blom A, Kafienah W, Hollander A
    Stem Cells Dev, 2014;23(5):541-54.  2014
  7. Suppression of cancer-initiating cells and selection of adipose-derived stem cells cultured on biomaterials having specific nanosegments.
    Authors: Kao T, Lee H, Higuchi A, Ling Q, Yu W, Chou Y, Wang P, Suresh Kumar S, Chang Y, Hung Chen Y, Chang Y, Chen D, Hsu S
    J Biomed Mater Res B Appl Biomater, 2013;102(3):463-76.  2013
  8. miR-125b Is an adhesion-regulated microRNA that protects mesenchymal stem cells from anoikis.
    Authors: Yu X, Cohen DM, Chen CS
    Stem Cells, 2012;30(5):956-64.  2012
  9. Perivascular mesenchymal progenitors in human fetal and adult liver.
    Authors: Gerlach J, Over P, Turner M, Thompson R, Foka H, Chen W, Peault B, Gridelli B, Schmelzer E
    Stem Cells Dev, 2012;21(18):3258-69.  2012
  10. Long-lasting inhibitory effects of fetal liver mesenchymal stem cells on T-lymphocyte proliferation.
    Authors: Giuliani M, Fleury M, Vernochet A, Ketroussi F, Clay D, Azzarone B, Lataillade JJ, Durrbach A
    2011;6(5):e19988.  2011
  11. Long-term culture following ES-like gene-induced reprogramming elicits an aggressive phenotype in mutated cholangiocellular carcinoma cells.
    Authors: Nagai K, Ishii H, Miyoshi N, Hoshino H, Saito T, Sato T, Tomimaru Y, Kobayashi S, Nagano H, Sekimoto M, Doki Y, Mori M
    Biochem. Biophys. Res. Commun., 2010;395(2):258-63.  2010

FAQs

  1. 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?

    • 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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StemXVivo Human Osteogenic Supplement, 12.5 mL
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