StemXVivo Osteogenic/Adipogenic Base Media

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CCM007
StemXVivo Osteogenic/Adipogenic Base Media
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StemXVivo Osteogenic/Adipogenic Base Media Summary

Kit Summary

Base media for the differentiation of MSCs into osteocytes or adipocytes. For use with StemXVivo® Osteogenic or Adipogenic Supplements.

Key Benefits

  • Supports human, mouse, and rat MSC differentiation
  • Developed and optimized using MSCs to reduce experimental variation
  • Tested for consistent MSC differentiation
 

 

Why Induce Osteogenic or Adipogenic Differentiation in MSCs with Defined Media?

Despite the well-characterized factors and protocols used to differentiate mesenchymal stem/stromal cells (MSCs) into osteocytes or adipocytes, differentiation efficiencies can vary depending on the quality of the MSC starting population and the reagents used to expand and differentiate MSCs.

Osteogenic/Adipogenic Base Media:

  • Supports reproducible human, mouse, and rat MSC differentiation.
  • Offers flexibility to evaluate novel cytokine and growth factor combinations to induce osteogenesis or adipogenesis.
  • Has been developed and optimized using MSCs.
  • Can be used with StemXVivo® Osteogenic Supplement (Catalog # CCM008 or CCM009) or StemXVivo® Adipogenic Supplement (Catalog # CCM011) to reduce variation in differentiation.
 

 

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/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media Components

Supplied in a 250 mL volume, this defined media contains high quality factors to drive MSC differentiation into osteocytes or adipocytes when used with additional differentiation factors.

  • Supplemented with sodium bicarbonate but does not contain antibiotics.

*This kit requires supplements (not included), such as StemXVivo® Osteogenic Supplement (Catalog # CCM008 or CCM009), StemXVivo® Adipogenic Supplement (Catalog # CCM011), or user-selected cytokines and growth factors.

 

Data Examples
Detection of Osteocalcin in Human MSCs-differentiated Osteocytes
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Detection of Osteocalcin in Human MSCs-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).

Detection of Osteopontin in Mouse MSC-differentiated Osteocytes
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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 # NL001; red) and the nuclei were counterstained with DAPI (blue).

Detection of Osteocalcin in Rat MSC-differentiated Osteocytes
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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 # NL007; red) and the nuclei were counterstained with DAPI (blue).

Detection of FABP4 in Human MSC-differentiated Adipocytes
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Detection of FABP4 in Human MSC-differentiated Adipocytes. Human MSCs were differentiated for 14 days using the Human/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007) and Human/Mouse/Rat StemXVivo® Adipogenic Supplement (Catalog # CCM011). Mature differentiated adipocytes were detected with a Goat Anti-Mouse FABP4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1443). The cells were stained with a NorthernLights 557-conjugated Donkey Anti-Mouse Secondary Antibody (Catalog # NL001; red) and the nuclei were counterstained with DAPI (blue).

Detection of Osteocalcin in Rat MSC-differentiated Osteocytes
View Larger Image

Detection of FABP4 in Mouse MSC-differentiated Adipocytes. Mouse MSCs were differentiated for 14 days using the Human/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007) and Human/Mouse/Rat StemXVivo® Adipogenic Supplement (Catalog # CCM011). Mature differentiated adipocytes were detected with a Goat Anti-Mouse FABP4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1443). The cells were stained with a NorthernLights 557-conjugated Donkey Anti-Mouse Secondary Antibody (Catalog # NL007; red) and the nuclei were counterstained with DAPI (blue).

Detection of Osteocalcin in Rat MSC-differentiated Osteocytes
View Larger Image

Detection of FABP4 in Rat MSC-differentiated Adipocytes. Rat MSCs were differentiated for 14 days using the Human/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007) and Human/Mouse/Rat StemXVivo® Adipogenic Supplement (Catalog # CCM011). Mature differentiated adipocytes were detected with a Goat Anti-Mouse FABP4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1443). The cells were stained with a NorthernLights 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 Mouse Rat

Product Datasheets

Data Example

StemXVivo® Osteogenic/Adipogenic Base Media

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, human, mouse, or rat MSCs are differentiated into adipocytes or osteocytes using the following in vitro differentiation procedure:

  • Culture multipotent cells of interest
  • Induce adipogenic or osteogenic differentiation using media supplements
  • Evaluate differentiation using a mature phenotype marker antibody and fluorescent ICC

For use with Human StemXVivo® Osteogenic Supplement (Catalog # CCM008), Mouse/Rat Osteogenic Supplement (Catalog # CCM009), or Human/Mouse/Rat StemXVivo® Adipogenic Supplement (Catalog # CCM011).

 

 

Reagents Provided

Reagents supplied in the Human/Mouse/Rat StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007):

  • 250 mL of StemXVivo® Osteogenic/Adipogenic Base Media

 

Other Supplies Required

Reagents

  • Human StemXVivo® Osteogenic Supplement (Catalog # CCM008) or Mouse/Rat Osteogenic Supplement (Catalog # CCM009)
  • Human/Mouse/Rat StemXVivo® Adipogenic Supplement (Catalog # CCM011)
  • Penicillin-Streptomycin-Glutamate (100X)

Materials

  • 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.

Osteogenic Differentiation

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

Osteogenic Differentiation

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

 

After 14-21 days, osteocytes can be harvested and analyzed.

Osteogenic Differentiation

Adipogenic Differentiation

 

Plate 2.1 x 104 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.

Culture cells to 100% confluency.

Subculturing Mesenchymal Stem Cells

Replace the medium in with Adipogenic Differentiation Medium to induce adipogenesis.

Subculturing Mesenchymal Stem Cells

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

 

After 14-21 days, adipocytes can be harvested and analyzed.

Subculturing Mesenchymal Stem Cells

Reagents Provided

Citations for StemXVivo Osteogenic/Adipogenic Base Media

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.

29 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. Cladophora glomerata methanolic extract promotes chondrogenic gene expression and cartilage phenotype differentiation in equine adipose-derived mesenchymal stromal stem cells affected by metabolic syndrome
    Authors: L Bourebaba, I Michalak, M Baouche, K Kucharczyk, K Marycz
    Stem Cell Res Ther, 2019;10(1):392.  2019
  3. Maintenance of stem cell viability and differentiation potential following cryopreservation within 3-dimensional hyaluronic acid hydrogels
    Authors: S Khetan, O Corey
    Cryobiology, 2019;0(0):.  2019
  4. 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
  5. Evaluation of Oxidative Stress and Mitophagy during Adipogenic Differentiation of Adipose-Derived Stem Cells Isolated from Equine Metabolic Syndrome (EMS) Horses
    Authors: K Marycz, C Weiss, A ?mieszek, K Kornicka
    Stem Cells Int, 2018;2018(0):5340756.  2018
  6. Mesenchymal Stem Cells Promote the Osteogenesis in Collagen-Induced Arthritic Mice through the Inhibition of TNF-?
    Authors: C Liu, H Zhang, X Tang, R Feng, G Yao, W Chen, W Li, J Liang, X Feng, L Sun
    Stem Cells Int, 2018;2018(0):4069032.  2018
  7. 1,25-Dihydroxyvitamin D suppresses M1 macrophages and promotes M2 differentiation at bone injury sites
    Authors: S Wasnik, CH Rundle, DJ Baylink, MS Yazdi, EE Carreon, Y Xu, X Qin, KW Lau, X Tang
    JCI Insight, 2018;3(17):.  2018
  8. 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
  9. Chemotherapy-induced niche perturbs hematopoietic reconstitution in B-cell acute lymphoblastic leukemia
    Authors: C Tang, MH Li, YL Chen, HY Sun, SL Liu, WW Zheng, MY Zhang, H Li, W Fu, WJ Zhang, AB Liang, ZH Tang, DL Hong, BS Zhou, CW Duan
    J. Exp. Clin. Cancer Res., 2018;37(1):204.  2018
  10. Long-term regeneration and remodeling of the pig esophagus after circumferential resection using a retrievable synthetic scaffold carrying autologous cells
    Authors: S La Frances, JM Aho, MR Barron, EW Blanco, S Soliman, L Kalenjian, AD Hanson, E Todorova, M Marsh, K Burnette, H DerSimonia, RD Odze, DA Wigle
    Sci Rep, 2018;8(1):4123.  2018
  11. Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods
    Authors: HR Lin, CW Heish, CH Liu, S Muduli, HF Li, A Higuchi, SS Kumar, AA Alarfaj, MA Munusamy, ST Hsu, DC Chen, G Benelli, K Murugan, NC Cheng, HC Wang, GJ Wu
    Sci Rep, 2017;7(0):40069.  2017
  12. Human chorionic villous mesenchymal stem/stromal cells modify the effects of oxidative stress on endothelial cell functions
    Authors: MH Abumaree, M Hakami, FM Abomaray, MA Alshabibi, B Kalionis, MA Al Jumah, AS AlAskar
    Placenta, 2017;0(0):.  2017
  13. An IGF1R-Dependent Pathway Drives Epicardial Adipose Tissue Formation After Myocardial Injury
    Authors: Kenneth R Chien
    Circulation, 2016;0(0):.  2016
  14. Macroautophagy and Selective Mitophagy Ameliorate Chondrogenic Differentiation Potential in Adipose Stem Cells of Equine Metabolic Syndrome: New Findings in the Field of Progenitor Cells Differentiation
    Authors: K Marycz, K Kornicka, J Grzesiak, A ?mieszek, J Sz?apka
    Oxid Med Cell Longev, 2016;2016(0):3718468.  2016
  15. Enhancement of Adipocyte Browning by Angiotensin II Type 1 Receptor Blockade
    PLoS ONE, 2016;11(12):e0167704.  2016
  16. Cat amniotic membrane multipotent cells are nontumorigenic and are safe for use in cell transplantation.
    Authors: Vidane A, Souza A, Sampaio R, Bressan F, Pieri N, Martins D, Meirelles F, Miglino M, Ambrosio C
    Stem Cells Cloning, 2014;7(0):71-8.  2014
  17. MCP/CCR2 signaling is essential for recruitment of mesenchymal progenitor cells during the early phase of fracture healing.
    Authors: Ishikawa, Masahiro, Ito, Hiromu, Kitaori, Toshiyuk, Murata, Koichi, Shibuya, Hideyuki, Furu, Moritosh, Yoshitomi, Hiroyuki, Fujii, Takayuki, Yamamoto, Koji, Matsuda, Shuichi
    PLoS ONE, 2014;9(8):e104954.  2014
  18. Purification of human adipose-derived stem cells from fat tissues using PLGA/silk screen hybrid membranes.
    Authors: Chen D, Chen L, Ling Q, Wu M, Wang C, Suresh Kumar S, Chang Y, Munusamy M, Alarfajj A, Wang H, Hsu S, Higuchi A
    Biomaterials, 2014;35(14):4278-87.  2014
  19. 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
  20. Label retention identifies a multipotent mesenchymal stem cell-like population in the postnatal thymus.
    Authors: Osada M, Singh V, Wu K, Sant'Angelo D, Pezzano M
    PLoS ONE, 2013;8(12):e83024.  2013
  21. Molecular characterization of prospectively isolated multipotent mesenchymal progenitors provides new insight into the cellular identity of mesenchymal stem cells in mouse bone marrow.
    Authors: Qian H, Badaloni A, Chiara F, Stjernberg J, Polisetti N, Nihlberg K, Consalez G, Sigvardsson M
    Mol Cell Biol, 2013;33(4):661-77.  2013
  22. Distinct fibroblast lineages determine dermal architecture in skin development and repair.
    Authors: Driskell R, Lichtenberger B, Hoste E, Kretzschmar K, Simons B, Charalambous M, Ferron S, Herault Y, Pavlovic G, Ferguson-Smith A, Watt F
    Nature, 2013;504(7479):277-81.  2013
  23. 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
  24. 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
  25. 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
  26. Lung-derived mesenchymal stromal cell post-transplantation survival, persistence, paracrine expression, and repair of elastase-injured lung.
    Authors: Hoffman AM, Paxson JA, Mazan MR, Davis AM, Tyagi S, Murthy S, Ingenito EP
    Stem Cells Dev., 2011;20(10):1779-92.  2011
  27. Mesenchymal stem cells stably transduced with a dominant-negative inhibitor of CCL2 greatly attenuate bleomycin-induced lung damage.
    Authors: Saito S, Nakayama T, Hashimoto N, Miyata Y, Egashira K, Nakao N, Nishiwaki S, Hasegawa M, Hasegawa Y, Naoe T
    Am. J. Pathol., 2011;179(3):1088-94.  2011
  28. 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
  29. 3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells.
    Authors: Wang W, Itaka K, Ohba S, Nishiyama N, Chung UI, Yamasaki Y, Kataoka K
    Biomaterials, 2009;30(14):2705-15.  2009

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