StemXVivo Human/Mouse Chondrogenic Supplement, 0.5 mL

Catalog # Availability Size / Price Qty
CCM006
Product Details
Procedure
Citations (16)
FAQs
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StemXVivo Human/Mouse Chondrogenic Supplement, 0.5 mL Summary

Kit Summary

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

Key Benefits

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

 

Why Induce Chondrogenesis in MSCs with a Defined Media Supplement?

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.

Human/Mouse StemXVivo® Chondrogenic Supplement:

  • Contains high quality differentiation factors to drive reproducible and efficient MSC chondrogenesis.
  • Is defined to reduce unwanted experimental variability.
  • Has been developed and optimized using human and mouse 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/Mouse Chondrogenic Supplement Components

This media supplement contains defined, high quality factors to drive MSC differentiation into chondrocytes.

  • This supplement requires media (not included), such as Human/Mouse/Rat StemXVivo® Chondrogenic Base Media (Catalog # CCM005) or equivalent.
  • The quantity of chondrogenic media supplement supplied is sufficient to make 50 mL of media for differentiation.

 

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

Detection of Aggrecan in a Human MSC-differentiated Chondrogenic Pellet Section
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Detection of Aggrecan in a Human MSC-differentiated Chondrogenic Pellet Section. Human MSCs cultured with Human/Mouse/Rat 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 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) and the nuclei were counterstained with DAPI.

Detection of Collagen II in a Mouse MSC-differentiated Chondrogenic Pellet Section
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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).

 

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

Product Datasheets

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, human or mouse 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/Rat StemXVivo® Chondrogenic Base Media (Catalog # CCM005).

 

 

Reagents Provided

Reagents supplied in the Human/Mouse StemXVivo® Chondrogenic Supplement (Catalog # CCM006):

  • 0.5 mL of StemXVivo® Chondrogenic Supplement

 

Other Supplies Required

Reagents

  • Human/Mouse/Rat StemXVivo® Chondrogenic Base Media (Catalog # CCM005)
  • 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

Reagents Provided

Citations for StemXVivo Human/Mouse Chondrogenic Supplement, 0.5 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.

16 Citations: Showing 1 - 10
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  1. Therapeutic Potential of Dental Pulp Stem Cells and Leukocyte- and Platelet-Rich Fibrin for Osteoarthritis
    Authors: M Lo Monaco, P Gervois, J Beaumont, P Clegg, A Bronckaers, JM Vandeweerd, I Lambrichts
    Cells, 2020;9(4):.  2020
  2. Comparison of Immunosuppressive and Angiogenic Properties of Human Amnion-Derived Mesenchymal Stem Cells between 2D and 3D Culture Systems
    Authors: V Miceli, M Pampalone, S Vella, AP Carreca, G Amico, PG Conaldi
    Stem Cells Int, 2019;2019(0):7486279.  2019
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. Mesenchymal stromal cells improve salivary function and reduce lymphocytic infiltrates in mice with Sjogren's-like disease.
    Authors: Khalili S, Liu Y, Kornete M, Roescher N, Kodama S, Peterson A, Piccirillo CA, Tran SD
    PLoS ONE, 2012;7(6):e38615.  2012
  14. 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
  15. 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
  16. Acute myeloid leukemia transforms the bone marrow niche into a leukemia-permissive microenvironment through exosome secretion.
    Authors: Kumar B, Garcia M, Weng L, Jung X, Murakami J, Hu X, McDonald T, Lin A, Kumar A, DiGiusto D, Stein A, Pullarkat V, Hui S, Carlesso N, Kuo Y, Bhatia R, Marcucci G, Chen C
    Leukemia, 0;32(3):575-587.  0

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).

  2. 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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