StemXVivo Chondrogenic Base Media Summary
Base media for the differentiation of MSCs into chondrocytes. For use with Human/Mouse and Rat StemXVivo® Chondrogenic Supplements.
- 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.
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
- 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.
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.
- Dominici, M. et al. (2006) Cytotherapy 8:315.
- Keating, A. (2012) Cell Stem Cell 10:709.
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).
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.
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).
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).
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
Reagents supplied in the Human/Mouse/Rat Chondrogenic Base Media (Catalog # CCM005):
- 50 mL of StemXVivo® Chondrogenic Base Media
- Human/Mouse StemXVivo® Chondrogenic Supplement (Catalog # CCM006) or Rat Chondrogenic Supplement (Catalog # CCM020)
- Penicillin-Streptomycin-Glutamate (100X)
- 15 mL centrifuge tubes
- Pipettes and pipette tips
- Serological pipettes
- 37 °C and 5% CO2 incubator
- 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.
Transfer 2.5 x 105 MSCs to a 15 mL conical tube.
Centrifuge and resuspend the cells in Chondrogenic Differentiation Medium.
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.
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.
Citations: Showing 1 - 10
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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;12(3):1074-1096. 2022
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;17(6):1376-1386. 2022
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;36(2):109352. 2021
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;10(1):4326. 2020
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;2019(0):9323864. 2019
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
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;2018(0):1310904. 2018
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
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;12(2):e0172430. 2017
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
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;12(1):e0166014. 2017
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;4(4):359-68. 2015
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
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;2014(0):468927. 2014
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;0(0):1-8. 2014
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
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
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;287(31):25795-807. 2012
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
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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