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StemXVivo Chondrogenic Base Media

Catalog #: CCM005
22 Citations Datasheet
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CCM005
Detection of Aggrecan in a Human MSC-differentiated Chondrogenic Pellet Section.
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Product Details
Procedure
Citations (22)
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StemXVivo Chondrogenic Base Media Summary

Kit Summary

Base media for the differentiation of MSCs into chondrocytes. For use with Human/Mouse and Rat StemXVivo® Chondrogenic Supplements.

Key Benefits

  • Defined formulation that reduces experimental variation
  • Supports induction of chondrogenesis in MSCs
  • Developed and optimized using MSCs

Why Induce Chondrogenesis in MSCs with Defined Media?

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

StemXVivo® Chondrogenic Base Media:

  • Is defined to support reproducible MSC chondrogenesis.
  • Offers flexibility to evaluate novel cytokine and growth factor combinations to induce chondrogenesis.
  • Has been developed and optimized using MSCs.
  • Can be used with StemXVivo® Human/Mouse or Rat Chondrogenic Supplements to reduce variation during chondrogenesis.

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® Chondrogenic Base Media

Supplied in a 50 mL volume, this media contains high quality factors to drive MSC differentiation into chondrocytes when used with additional differentiation factors.

  • Supplemented with sodium bicarbonate but does not contain antibiotics.

*This medium requires supplements (not included), such as Human/Mouse StemXVivo® Chondrogenic Supplement (Catalog # CCM006), Rat StemXVivo® Chondrogenic Supplement (Catalog # CCM020), or user-defined cytokines and growth factors to induce chondrogenesis.

Precautions

This product contains human transferrin. The transferrin was purified from donor plasma and tested at the donor level using an FDA licensed method and found to be non-reactive for anti-HIV-1/2 and Hepatitis B surface antigen. 

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.

Specifications

Source
N/A
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

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Scientific Data

Immunocytochemistry Detection of Aggrecan in a Human MSC-differentiated Chondrogenic Pellet Section. View Larger

Detection of Aggrecan in a Human MSC-differentiated Chondrogenic Pellet Section. Human MSCs were cultured with StemXVivo®Chondrogenic Base Media (Catalog # CCM005) and StemXVivo®Chondrogenic Supplement (Catalog # CCM006) and the resulting chondrogenic pellet was cryosectioned. Chondrocyte differentiation was verified using a Goat Anti-Human Aggrecan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1220). 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).

Cell Morphology MSCs Differentiated into Chondrocytes form Characteristic Cell Pellets. View Larger

MSCs Differentiated into Chondrocytes form Characteristic Cell Pellets. MSCs Differentiated into Chondrocytes form Characteristic Cell Pellets.Human MSCs cultured with StemXVivo Chondrogenic Base Media (Catalog # CCM005) and StemXVivo Chondrogenic Supplement (Catalog # CCM006) formed a chondrogenic pellet (ball) imaged here at day 21 of culture.

Immunocytochemistry Detection of Collagen II in a Mouse MSC-differentiated Chondrogenic Pellet Section. View Larger

Detection of Collagen II in a Mouse MSC-differentiated Chondrogenic Pellet Section. Mouse MSCs were cultured for 21 days using the Human/Mouse StemXVivo®Chondrogenic Base Media (Catalog # ;CCM005) and Human/Mouse StemXVivo®Chondrogenic Supplement (Catalog # CCM006) and the resulting chondrogenic pellet was cryosectioned. Chondrocyte differentiation was verified using a Sheep Anti-Mouse Collagen II Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3615). The cells were stained using a NorthernLights™557-conjugated Donkey Anti-Sheep Secondary Antibody (Catalog # NL010; red) and the nuclei were counterstained with DAPI (blue).

Immunocytochemistry Detection of Aggrecan in a Rat MSC-differentiated Chondrogenic Pellet Section. View Larger

Detection of Aggrecan in a Rat MSC-differentiated Chondrogenic Pellet Section. Rat MSCs were cultured for 21 days using the Human/Mouse StemXVivo®Chondrogenic Base Media (Catalog # CCM005) and Rat StemXVivo®Chondrogenic Supplement (Catalog # CCM020) and the resulting chondrogenic pellet was cryosectioned. Chondrocyte differentiation was verified using a Goat Anti-Human Aggrecan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1220). 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).

Assay Procedure

Refer to the product datasheet for complete product details.

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

  • Culture multipotent cells of interest
  • Induce chondrogenic differentiation using a media supplement
  • Evaluate differentiation using a mature phenotype marker antibody and fluorescent ICC

For use with Human/Mouse StemXVivo® Chondrogenic Supplement (Catalog # CCM006) or Rat StemXVivo® Chondrogenic Supplement (Catalog # CCM020).

CCM005 Protocol Graphic Summary
View Graphic Procedure

 

Reagents Provided

Reagents supplied in the Human/Mouse/Rat Chondrogenic Base Media (Catalog # CCM005):

  • 50 mL of StemXVivo® Chondrogenic Base Media

Other Supplies Required

Reagents

  • Human/Mouse StemXVivo® Chondrogenic Supplement (Catalog # CCM006) or Rat Chondrogenic Supplement (Catalog # CCM020)
  • Penicillin-Streptomycin-Glutamate (100X)

Materials

  • MSCs
  • 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.

Transfer 2.5 x 105 MSCs to a 15 mL conical tube.

Centrifuge and resuspend the cells in Chondrogenic Differentiation Medium.

Protocol for Human CD4+ T Cell Enrichment Column

Centrifuge the cells but do not remove the medium.

Every 2-3 days, replace with fresh Chondrogenic Differentiation Medium.

After 14-21 days, the chondrogenic pellet can be harvested and analyzed.

Protocol for Human CD4+ T Cell Enrichment Column

Citations for StemXVivo Chondrogenic 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.

22 Citations: Showing 1 - 10
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  1. 3D culture of mesenchymal stem cells from the yolk sac to generate intestinal organoid
    Authors: Motta, LCB;Pereira, VM;Pinto, PAF;Mançanares, CAF;Pieri, NCG;de Oliveira, VC;Fantinato-Neto, P;Ambrósio, CE;
    Theriogenology  2023-06-16
  2. Periosteal skeletal stem cells can migrate into the bone marrow and support hematopoiesis after injury
    Authors: Marchand, T;Akinnola, KE;Takeishi, S;Maryanovich, M;Pinho, S;Saint-Vanne, J;Birbrair, A;Lamy, T;Tarte, K;Frenette, PS;Gritsman, K;
    bioRxiv : the preprint server for biology  2023-01-13
  3. Effects of amyloid precursor protein peptide APP96-110, alone or with human mesenchymal stromal cells, on recovery after spinal cord injury
    Authors: SI Hodgetts, SJ Lovett, D Baron-Heer, A Fogliani, M Sturm, C Van den He, AR Harvey
    Neural regeneration research, 2022-06-01;17(6):1376-1386.  2022-06-01
  4. A FoxA2+ long-term stem cell population is necessary for growth plate cartilage regeneration after injury
    Authors: S Muruganand, R Pierce, DA Teguh, RF Perez, N Bell, B Nguyen, K Hohl, BD Snyder, MW Grinstaff, H Alberico, D Woods, Y Kong, C Sima, S Bhagat, K Ho, V Rosen, L Gamer, AM Ionescu
    Nature Communications, 2022-05-06;13(1):2515.  2022-05-06
  5. Single-cell RNA landscape of the osteoimmunology microenvironment in periodontitis
    Authors: Y Chen, H Wang, Q Yang, W Zhao, Y Chen, Q Ni, W Li, J Shi, W Zhang, L Li, Y Xu, H Zhang, D Miao, L Xing, W Sun
    Theranostics, 2022-01-01;12(3):1074-1096.  2022-01-01
  6. Regional specialization and fate specification of bone stromal cells in skeletal development
    Authors: KK Sivaraj, HW Jeong, B Dharmaling, D Zeuschner, S Adams, M Potente, RH Adams
    Cell Reports, 2021-07-13;36(2):109352.  2021-07-13
  7. Transplantation of a 3D-printed tracheal graft combined with iPS cell-derived MSCs and chondrocytes
    Authors: IG Kim, SA Park, SH Lee, JS Choi, H Cho, SJ Lee, YW Kwon, SK Kwon
    Sci Rep, 2020-03-09;10(1):4326.  2020-03-09
  8. 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, 2019-12-17;10(1):392.  2019-12-17
  9. Metabolic Phenotyping of Adipose-Derived Stem Cells Reveals a Unique Signature and Intrinsic Differences between Fat Pads
    Authors: C Lefevre, B Panthu, D Naville, S Guibert, C Pinteur, B Elena-Herr, H Vidal, GJP Rautureau, A Mey
    Stem Cells Int, 2019-05-14;2019(0):9323864.  2019-05-14
  10. Efficient Nonviral Transfection of Human Bone Marrow Mesenchymal Stromal Cells Shown Using Placental Growth Factor Overexpression
    Authors: WY Cheung, O Hovey, JM Gobin, G Muradia, J Mehic, C Westwood, JR Lavoie
    Stem Cells Int, 2018-12-24;2018(0):1310904.  2018-12-24
  11. 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., 2018-08-29;37(1):204.  2018-08-29
  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, 2017-05-04;0(0):.  2017-05-04
  13. Mesenchymal stromal cells (MSCs) induce ex vivo proliferation and erythroid commitment of cord blood haematopoietic stem cells (CB-CD34+ cells)
    Authors: S Perucca, A Di Palma, PP Piccaluga, C Gemelli, E Zoratti, G Bassi, E Giacopuzzi, A Lojacono, G Borsani, E Tagliafico, MT Scupoli, S Bernardi, C Zanaglio, F Cattina, V Cancelli, M Malagola, M Krampera, M Marini, C Almici, S Ferrari, D Russo
    PLoS ONE, 2017-02-23;12(2):e0172430.  2017-02-23
  14. Mesenchymal Cell Reprogramming in Experimental MPLW515L Mouse Model of Myelofibrosis
    Authors: Y Han, L Yue, M Wei, X Ren, Z Shao, L Zhang, RL Levine, PK Epling-Bur
    PLoS ONE, 2017-01-30;12(1):e0166014.  2017-01-30
  15. Peripheral blood-derived mesenchymal stem cells: candidate cells responsible for healing critical-sized calvarial bone defects.
    Authors: Li S, Huang K, Wu J, Hu M, Sanyal M, Hu M, Longaker M, Lorenz H
    Stem Cells Transl Med, 2015-03-05;4(4):359-68.  2015-03-05
  16. Bone marrow-derived multipotent stromal cells attenuate inflammation in obliterative airway disease in mouse tracheal allografts.
    Authors: Casey A, Dirks F, Liang O, Harrach H, Schuette-Nuetgen K, Leeman K, Kim C, Gerard C, Subramaniam M
    Stem Cells Int, 2014-09-10;2014(0):468927.  2014-09-10
  17. Improved quality of cartilage repair by bone marrow mesenchymal stem cells for treatment of an osteochondral defect in a cynomolgus macaque model.
    Authors: Araki S, Imai S, Ishigaki H, Mimura T, Nishizawa K, Ueba H, Kumagai K, Kubo M, Mori K, Ogasawara K, Matsusue Y
    Acta Orthop, 2014-09-01;0(0):1-8.  2014-09-01
  18. 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, 2014-08-27;7(0):71-8.  2014-08-27
  19. 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, 2012-11-26;33(4):661-77.  2012-11-26
  20. 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, 2012-10-16;21(18):3258-69.  2012-10-16
  21. Primary Mesenchymal Stem and Progenitor Cells from Bone Marrow Lack Expression of CD44 Protein.
    Authors: Qian H, Le Blanc K, Sigvardsson M
    J. Biol. Chem., 2012-05-31;287(31):25795-807.  2012-05-31
  22. 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
    2011-05-19;6(5):e19988.  2011-05-19

FAQs

  1. Are there any experimental tips/hints for successful chondrogenic differentiation of mesenchymal stem cells?

    • The following tips/hints are useful for chondrogenic differentiation:

      a) The mesenchymal stem cells (MSCs) should not be from a late passage (passage 8 or less), b) if using the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006) or the StemXVivo® Chondrogenic Supplement (Catalog # CCM006), use the starting MSC cell number that is indicated in the protocol, c) Early during chondrogenic differentiation a pellet should form. As differentiation progresses, the pellet will grow and take up a ball-like appearance. d) The pellet should not attach to the tube, therefore care should be taken to not dislodge it while changing media.

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