Human Mesenchymal Stem Cell Functional Identification Kit
Catalog # SC006 | R&D Systems, Inc. a Bio-Techne Brand
Key Product Details
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, human MSC multipotency is verified using the following in vitro differentiation procedure:
- Culture multipotent cells of interest
- Induce adipocyte, chondrocyte, and osteocyte differentiation using media supplements
- Evaluate differentiation using mature phenotype marker antibodies and fluorescent ICC
Reagents supplied in the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006):
- Adipogenic Supplement
- Chondrogenic Supplement
- Osteogenic Supplement
- ITS Supplement
- Adipocyte marker: Goat Anti-Mouse FABP4 Antigen-affinity Purified Polyclonal Antibody
- Chondrocyte marker: Goat Anti-Human Aggrecan Antigen-affinity Purified Polyclonal Antibody
- Osteocyte marker: Mouse Anti-Human Osteocalcin Antigen-affinity Purified Monoclonal Antibody
Note: The quantity of each media supplement in this kit is sufficient to make 50 mL of media for differentiation. 50 mL can be used for 16 wells of a 24-well plate for osteogenic and adipogenic lineages and 10 chondrocyte pellets.
Reagents
- StemXVivo® Osteogenic/Adipogenic Base Media (Catalog # CCM007 or equivalent)
- D-MEM/F-12 (1X)
- Phosphate Buffered Saline (PBS)
- Penicillin-Streptomycin-Glutamate (100X)
- 4% Paraformaldehyde in PBS
- 1% BSA in PBS
- Triton® X-100
- 10% normal donkey serum
- Fibronectin (optional; Human Fibronectin, Catalog # 1918-FN, Bovine Fibronectin, Catalog # 1030-FN, or equivalent)
- Mounting medium (Catalog # CTS011 or equivalent)
- NorthernLightsTM 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001 or equivalent)
- Deionized or distilled water
Materials
- Human MSCs
- 24-well culture plates
- 12 mm coverslips (Carolina Biologicals, Catalog # 633009 or equivalent)
- 15 mL centrifuge tubes
- Pipettes and pipette tips
- Serological pipettes
- Glass slides
- Fine pointed curved forceps
- Liquid barrier pen
Equipment
- 37 °C and 5% CO2 incubator
- Centrifuge
- Hemocytometer
- Inverted microscope
- 2 °C to 8 °C refrigerator
- 37 °C water bath
- Fluorescence microscope
- Cryostat
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.
Adipogenic Differentiation
Plate 2.1 x 104 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.
Culture cells to 100% confluency.

Replace the medium with Adipogenic Differentiation Medium to induce adipogenesis.

Every 2-3 days, replace with fresh Adipogenic Differentiation Medium.
After 14-21 days, adipocytes can be fixed.
ICC detection of FABP4.

Osteogenic Differentiation
Plate 4.2 x 103 MSCs/cm2 in StemXVivo® Osteogenic/Adipogenic Base Media.
Culture cells to 50-70% confluency.

Replace the medium with Osteogenic Differentiation Medium to induce osteogenesis.

Every 3-4 days, replace with fresh Osteogenic Differentiation Medium.
After 14-21 days, osteocytes can be fixed.
ICC detection of Osteocalcin.

Chondrogenic Differentiation
Transfer 2.5 x 104 MSCs to a 15 mL conical tube.
Centrifuge and resuspend the cells in Chondrogenic Differentiation Media.
Centrifuge the cells but do not remove the medium.

Every 2-3 days, replace with fresh Chondrogenic Differentiation Media.
After 14-21 days, the chondrogenic pellet can be fixed.

Cryosection the chondrogenic pellet.
ICC detection of Aggrecan.

Citations for Human Mesenchymal Stem Cell Functional Identification Kit (63)
Citations are publications that use Bio-Techne products. Selected citations for Human Mesenchymal Stem Cell Functional Identification Kit include:
-
Species: Human
Sample Types: Whole Cells
Wu et al. (2023-05-12), Directed differentiation of human iPSCs into mesenchymal lineages by optogenetic control of TGF-? signaling Cell reports
PMID: 37178118 -
Species: Human, Rat
Sample Types: Whole Cells
NC Brembilla et al. (2023-03-22), Adipose-Derived Stromal Cells within a Gelatin Matrix Acquire Enhanced Regenerative and Angiogenic Properties: A Pre-Clinical Study for Application to Chronic Wounds Biomedicines, 2023-011(3).
PMID: 36979966 -
Species: Human
Sample Types: Whole Cells
C Río et al. (2023-03-18), Mesenchymal Stem Cells from COPD Patients Are Capable of Restoring Elastase-Induced Emphysema in a Murine Experimental Model International Journal of Molecular Sciences, 2023-024(6).
PMID: 36982887 -
Species: Human
Sample Types: Whole Cells
S Saber et al. (2023-03-17), Effect of Different Sealers on the Cytocompatibility and Osteogenic Potential of Human Periodontal Ligament Stem Cells: An In Vitro Study Journal of Clinical Medicine, 2023-012(6).
PMID: 36983344 -
Species: Human
Sample Types: Whole Cell
V Vitus et al. (2022-12-15), Carbonised Human Hair Incorporated in Agar/KGM Bioscaffold for Tissue Engineering Application: Fabrication and Characterisation Polymers, 2022-114(24).
PMID: 36559856 -
E Anitua et al. (2022-03-02), Composite alginate-gelatin hydrogels incorporating PRGF enhance human dental pulp cell adhesion, chemotaxis and proliferation International journal of pharmaceutics, 2022-0617(0):121631.
PMID: 35247496 -
Species: Human
Sample Types: Whole Cells
N Polisetti et al. (2022-03-02), Efficient Isolation and Functional Characterization of Niche Cells from Human Corneal Limbus International Journal of Molecular Sciences, 2022-023(5).
PMID: 35269891 -
Species: Human
Sample Types: Whole Cells
S El Harane et al. (2022-02-11), Adipose-derived stem cell spheroids are superior to single-cell suspensions to improve fat autograft long-term survival Journal of Cellular and Molecular Medicine, 2022-00(0).
PMID: 35150064 -
Species: Human
Sample Types: Whole Cells
S Zheng et al. (2022-01-01), A Robust and Highly Efficient Approach for Isolation of Mesenchymal Stem Cells From Wharton's Jelly for Tissue Repair Cell Transplantation, 2022-031(0):9636897221084.
PMID: 35313748 -
Species: Human
Sample Types: Whole Cells
Q Heydt et al. (2021-09-17), Adipocytes disrupt the translational programme of acute lymphoblastic leukaemia to favour tumour survival and persistence Nature Communications, 2021-012(1):5507.
PMID: 34535653 -
Species: Human
Sample Types: Whole Cells
A Mik?osz et al. (2021-06-16), Does TBC1D4 (AS160) or TBC1D1 Deficiency Affect the Expression of Fatty Acid Handling Proteins in the Adipocytes Differentiated from Human Adipose-Derived Mesenchymal Stem Cells (ADMSCs) Obtained from Subcutaneous and Visceral Fat Depots? Cells, 2021-010(6).
PMID: 34208471 -
Species: Xenopus
Sample Types: Whole Cells
R Otsuka-Yam et al. (2021-04-08), Isolation and characterization of bone marrow-derived mesenchymal stem cells in Xenopus laevis Stem Cell Research, 2021-053(0):102341.
PMID: 33892293 -
Species: Human
Sample Types: Whole Cells
J Jazowiecka et al. (2021-03-19), Myxoma Virus Expressing LIGHT (TNFSF14) Pre-Loaded into Adipose-Derived Mesenchymal Stem Cells Is Effective Treatment for Murine Pancreatic Adenocarcinoma Cancers, 2021-013(6).
PMID: 33808692 -
Species: Human
Sample Types: Whole Cells
EM Wilfong et al. (2020-10-06), Proinflammatory cytokines and ARDS pulmonary edema fluid induce CD40 on human mesenchymal stromal cells-A potential mechanism for immune modulation PLoS ONE, 2020-115(10):e0240319.
PMID: 33021986 -
Species: Human
Sample Types: Cell Culture Supernates
S Liu et al. (2020-07-03), Assessment and Comparison of the Efficacy of Methotrexate, Prednisolone, Adalimumab, and Tocilizumab on Multipotency of Mesenchymal Stem Cells Front Pharmacol, 2020-011(0):1004.
PMID: 32719606 -
Species: Human
Sample Types: Whole Cells
J Wu et al. (2020-06-16), Immunity-and-matrix-regulatory cells derived from human embryonic stem cells safely and effectively treat mouse lung injury and fibrosis Cell Res., 2020-00(0).
PMID: 32546764 -
Species: Human
Sample Types: Cell Culture Supernates
J Xu et al. (2020-03-19), Chemical-defined medium supporting the expansion of human mesenchymal stem cells Stem Cell Res Ther, 2020-011(1):125.
PMID: 32192530 -
Species: Human
Sample Types: Whole Cells
AI Marusina et al. (2019-11-23), Tunable hydrogels for mesenchymal stem cell delivery: integrin-induced transcriptome alterations and hydrogel optimization for human wound healing Stem Cells, 2019-10(0).
PMID: 31648388 -
Species: Human
Sample Types: Whole Cells
T Suzuki et al. (2019-11-18), Improving the viability of tissue-resident stem cells using an organ-preservation solution FEBS Open Bio, 2019-10(0).
PMID: 31642604 -
Species: Human
Sample Types: Whole Cells
TM Campbell et al. (2019-04-26), Tendon contains more stem cells than bone at the rotator cuff repair site J Shoulder Elbow Surg, 2019-00(0).
PMID: 31036422 -
Species: Human
Sample Types: Whole Cells
Y Lin et al. (2019-01-24), Combination of polyetherketoneketone scaffold and human mesenchymal stem cells from temporomandibular joint synovial fluid enhances bone regeneration Sci Rep, 2019-09(1):472.
PMID: 30679553 -
Species: Human
Sample Types: Whole Cells
TS Winston et al. (2019-01-15), Serum-Free Manufacturing of Mesenchymal Stem Cell Tissue Rings Using Human-Induced Pluripotent Stem Cells Stem Cells Int, 2019-02019(0):5654324.
PMID: 30766604 -
Species: Human
Sample Types: Whole Cells
J Ma et al. (2019-01-11), Comparative analysis of mesenchymal stem cells derived from amniotic membrane, umbilical cord, and chorionic plate under serum-free condition Stem Cell Res Ther, 2019-010(1):19.
PMID: 30635045 -
Species: Human
Sample Types: Whole Cells
Applications: BioassayBaulier E et al. (2018-11-28), Generation of a human Ocular Albinism type 1 iPSC line, SEIi001-A, with a mutation in GPR143. Stem Cell Res, 2018-133(0):274-277.
PMID: 30513407 -
Species: Human
Sample Types: Whole Cells
Applications: ICCE Toyoda et al. (2018-11-20), Multilineage-differentiating stress-enduring (Muse)-like cells exist in synovial tissue Regen Ther, 2018-110(0):17-26.
PMID: 30525067 -
Species: Human
Sample Types: Whole Cells
MH Abumaree et al. (2018-04-12), Characterization of the interaction between human decidua parietalis mesenchymal stem/stromal cells and natural killer cells Stem Cell Res Ther, 2018-09(1):102.
PMID: 29650045 -
Species: Human
Sample Types: Whole Cells
K Fujisawa et al. (2018-04-06), Evaluation of the effects of ascorbic acid on metabolism of human mesenchymal stem cells Stem Cell Res Ther, 2018-09(1):93.
PMID: 29625581 -
Species: Human
Sample Types: dental pulp cells
K Nakajima et al. (2018-03-11), Comparison of the bone regeneration ability between stem cells from human exfoliated deciduous teeth, human dental pulp stem cells and human bone marrow mesenchymal stem cells Biochem. Biophys. Res. Commun., 2018-0497(3):876-882.
PMID: 29477844 -
Species: Human
Sample Types: Whole Cells
H Wang et al. (2018-03-09), Comparative characterization of SHED and DPSCs during extended cultivation in�vitro Mol Med Rep, 2018-00(0).
PMID: 29532869 -
Species: Human
Sample Types: Whole Cells
S Cao et al. (2018-02-19), Nano-loaded human umbilical cord mesenchymal stem cells as targeted carriers of doxorubicin for breast cancer therapy Artif Cells Nanomed Biotechnol, 2018-00(0):1-11.
PMID: 29457930 -
Species: Human
Sample Types: Whole Cells
L Mezzanotte et al. (2017-12-01), Optimized Longitudinal Monitoring of Stem Cell Grafts in Mouse Brain Using a Novel Bioluminescent/Near Infrared Fluorescent Fusion Reporter Cell Transplant, 2017-126(12):1878-1889.
PMID: 29390874 -
Species: Human
Sample Types: Whole Cells
C Arianna et al. (2017-07-26), Rapid Rapamycin-Only Induced Osteogenic Differentiation of Blood-Derived Stem Cells and Their Adhesion to Natural and Artificial Scaffolds Stem Cells Int, 2017-02017(0):2976541.
PMID: 28814956 -
Species: Human
Sample Types: Whole Cells
H Takigawa et al. (2017-04-20), Mesenchymal Stem Cells Induce Epithelial to Mesenchymal Transition in Colon Cancer Cells through Direct Cell-to-Cell Contact Neoplasia, 2017-019(5):429-438.
PMID: 28433772 -
Species: Human
Sample Types: Whole Cells
K Tatebayash et al. (2017-04-19), Identification of multipotent stem cells in human brain tissue following stroke Stem Cells Dev, 2017-00(0).
PMID: 28323540 -
Species: Human
Sample Types: Whole Cells
V Zarrinpour et al. (2017-02-01), Expression pattern of neurotrophins and their receptors during neuronal differentiation of adipose-derived stem cells in simulated microgravity condition Iran J Basic Med Sci, 2017-020(2):178-186.
PMID: 28293395 -
Species: Human
Sample Types: Whole Cells
SW Lee et al. (2016-11-24), Are Adipose-Derived Stem Cells From Liver Falciform Ligaments Another Possible Source of Mesenchymal Stem Cells? Cell Transplant, 2016-126(5):855-866.
PMID: 27938473 -
Species: Human
Sample Types: Whole Cells
(2016-11-15), Human endometrial mesenchymal stem cells exhibit intrinsic anti-tumor properties on human epithelial ovarian cancer cells Sci Rep, 2016-16(0):37019.
PMID: 27845405 -
Species: Human
Sample Types: Whole Cells
Euiseong Kim (2016-10-12), In vitro characterization of human dental pulp stem cells isolated by three different methods Restor Dent Endod, 2016-141(4):283-295.
PMID: 27847750 -
Species: Human
Sample Types: Whole Cells
(2016-09-26), Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells Stem Cells Int, 2016-02016(0):5457132.
PMID: 27746820 -
Species: Human
Sample Types: Whole Cells
(2016-09-21), TLR3 preconditioning enhances the therapeutic efficacy of umbilical cord mesenchymal stem cells in TNBS-induced colitis via the TLR3-Jagged-1-Notch-1 pathway Mucosal Immunol, 2016-00(0).
PMID: 27649928 -
Species: Equine
Sample Types: Whole Cells
(2016-09-14), Equine metabolic syndrome impairs adipose stem cells osteogenic differentiation by predominance of autophagy over selective mitophagy J Cell Mol Med, 2016-00(0).
PMID: 27629697 -
Species: Human
Sample Types: Whole Cells
(2016-08-11), Exendin-4 enhances the differentiation of Wharton's jelly mesenchymal stem cells into insulin-producing cells through activation of various ?-cell markers Stem Cell Res Ther, 2016-07(1):108.
PMID: 27515427 -
Species: Human
Sample Types: Whole Cells
(2016-07-22), Transcriptome sequencing wide functional analysis of human mesenchymal stem cells in response to TLR4 ligand Sci Rep, 2016-06(0):30311.
PMID: 27444640 -
Species: Human
Sample Types: Whole Cells
Applications: Bioassay(2016-07-14), Human Cardiac Mesenchymal Stromal Cells with CD105+CD34- Phenotype Enhance the Function of Post-Infarction Heart in Mice PLoS ONE, 2016-011(7):e0158745.
PMID: 27415778 -
Species: Human
Sample Types: Whole Cells
Applications: Flow CytometryMatsuoka Y et al. (2015-05-01), Prospectively Isolated Human Bone Marrow Cell-Derived MSCs Support Primitive Human CD34-Negative Hematopoietic Stem Cells. Stem Cells, 2015-033(5):1554-65.
PMID: 25537923 -
Species: Human
Sample Types: Whole Cells
Manotham et al. (2015-03-26), Generation of CCR5-defective CD34 cells from ZFN-driven stop codon-integrated mesenchymal stem cell clones. J Biomed Sci, 2015-022(0):25.
PMID: 25890320 -
Species: Human
Sample Types: Whole Cells
Kim J et al. (2015-01-14), Umbilical cord mesenchymal stromal cells affected by gestational diabetes mellitus display premature aging and mitochondrial dysfunction. Stem Cells Dev, 2015-024(5):575-86.
PMID: 25437179 -
Species: Human
Sample Types: Whole Cells
Dossena M et al. (2014-11-13), Human adipose-derived mesenchymal stem cells as a new model of spinal and bulbar muscular atrophy. PLoS ONE, 2014-19(11):e112746.
PMID: 25392924 -
Species: Human
Sample Types: Whole Cells
Marfia G et al. (2014-10-29), Gene expression profile analysis of human mesenchymal stem cells from herniated and degenerated intervertebral discs reveals different expression of osteopontin. Stem Cells Dev, 2014-124(3):320-8.
PMID: 25203751 -
Species: Human
Sample Types: Whole Cells
Schade A et al. (2014-03-31), Magnetic Nanoparticle Based Nonviral MicroRNA Delivery into Freshly Isolated CD105(+) hMSCs. Stem Cells Int, 2014-02014(0):197154.
PMID: 24799915 -
Species: Human
Sample Types: Whole Cells
Dregalla et al. (2014-01-16), Amide-type local anesthetics and human mesenchymal stem cells: clinical implications for stem cell therapy. Stem Cells Transl Med, 2014-03(3):365-74.
PMID: 24436443 -
Species: Human
Sample Types: Whole Cells
Lehmann T et al. (2013-11-21), Coculture of human nucleus pulposus cells with multipotent mesenchymal stromal cells from human bone marrow reveals formation of tunnelling nanotubes. Mol Med Rep, 2013-19(2):574-82.
PMID: 24271232 -
Species: Human
Sample Types: Whole Cells
Gertow K et al. (2013-06-04), WNT3A promotes hematopoietic or mesenchymal differentiation from hESCs depending on the time of exposure. Stem Cell Reports, 2013-01(1):53-65.
PMID: 24052942 -
Species: Human
Sample Types: Whole Cells
Reinhardt P et al. (2013-03-22), Derivation and expansion using only small molecules of human neural progenitors for neurodegenerative disease modeling. PLoS ONE, 2013-08(3):e59252.
PMID: 23533608 -
Species: Human
Sample Types: Whole Cells
Raynaud C et al. (2013-01-16), Human embryonic stem cell derived mesenchymal progenitors express cardiac markers but do not form contractile cardiomyocytes. PLoS ONE, 2013-08(1):e54524.
PMID: 23342164 -
Species: Porcine
Sample Types: Whole Cells
Zhu X et al. (2013-01-01), Mesenchymal stem cells and endothelial progenitor cells decrease renal injury in experimental swine renal artery stenosis through different mechanisms. Stem Cells, 2013-031(1):117-25.
PMID: 23097349 -
Species: Human
Sample Types: Whole Cells
Applications: ICCSandt C et al. (2012-04-13), Identification of spectral modifications occurring during reprogramming of somatic cells. PLoS ONE, 2012-07(4):e30743.
PMID: 22514597 -
Species: Human
Sample Types: Whole Cells
Griscelli F et al. (2012-03-13), Malignant germ cell-like tumors, expressing Ki-1 antigen (CD30), are revealed during in vivo differentiation of partially reprogrammed human-induced pluripotent stem cells. Am. J. Pathol., 2012-0180(5):2084-96.
PMID: 22425713 -
Species: Human
Sample Types: Whole Cells
Tran NT et al. (2011-09-27), Efficient differentiation of human pluripotent stem cells into mesenchymal stem cells by modulating intracellular signaling pathways in a feeder/serum-free system. Stem Cells Dev., 2011-021(7):1165-75.
PMID: 21793661 -
Species: Human
Sample Types: Whole Cells
Roobrouck VD et al. (2011-05-01), Differentiation potential of human postnatal mesenchymal stem cells, mesoangioblasts, and multipotent adult progenitor cells reflected in their transcriptome and partially influenced by the culture conditions. Stem Cells, 2011-029(5):871-82.
PMID: 21433224 -
Species: Human
Sample Types: Whole Cells
Harvanova D et al. (2011-01-01), Isolation and characterization of synovial mesenchymal stem cells. Folia Biol. (Praha), 2011-057(3):119-24.
PMID: 21888835 -
Species: Human
Sample Types: Whole Cells
Fujino N et al. (2010-11-15), Isolation of alveolar epithelial type II progenitor cells from adult human lungs. Lab. Invest., 2010-191(0):363-78.
PMID: 21079581 -
Species: Human
Sample Types: Whole Cells
Han J et al. (2009-08-15), Collection and culture of alveolar bone marrow multipotent mesenchymal stromal cells from older individuals. J. Cell. Biochem., 2009-0107(6):1198-204.
PMID: 19507174
There are no citations that match your criteria.
Customer Reviews for Human Mesenchymal Stem Cell Functional Identification Kit (1)
Customer Images
-
Verified Customer | Posted 10/17/2016The MSC’s were generated from induced pluripotent stem cells and then tested using your kit for its potential to differentiate along the adipogenic and chondrogenic lineages.
There are no reviews that match your criteria.
FAQs for Human Mesenchymal Stem Cell Functional Identification Kit
-
Can the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006) be used with non-human primate mesenchymal stem cells?
It is likely that the antibodies included in the kit are cross-reactive to other primates. The supplements included in the kit are not intended to be species-specific. However, the kit has not been tested with primate mesenchymal stem cells
-
For the Human Mesenchymal Stem Cell Functional Identification Kit (Catalog # SC006), how can induction of differentiation be monitored?
For adipogenic differentiation, the appearance of vacuoles in cells after 5-7 days is a sign of differentiation and can be monitored by microscopic examination of the cells. For osteogenic differentiation, the beginning of cell detachment after about 14 days is a sign of differentiation. Cell detachment should be monitored in this case. For chondrogenic differentiation, there isn't an exact marker to look for other than fixing and staining the frozen pellet between differentiation days 14 - 21. The exact choice of time may take some empirical testing.
-
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).
-
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.