Dear Rajesh Singh,
I have been using StemPro® Chondrogenesis Differentiation Kit , and I worked for me!
If you have any other question please feel free to ask,
Best regards
MSC's usually show CD 10+, CD13+, CD 44+ , CD 73+ phenotype but they dont express CD 31 or 45. Surface marker analysis alongwith totipotency studies could help. Osteogenic differentiation of MSCs can be triggered by exposure to specific culture supplements (eg. dexamethasone, ascorbate-2-phosphate etc.) Adipogenic differentiation can also be induced through the use of induction medium containing dexamethasone, indomethacin, isobutylmethylxanthine, and insulin, but these cultures require a higher seeding density. Osteogenic differentiation of hMSCs can be triggered by exposure to specific culture supplements including dexamethasone, ascorbate-2-phosphate, and beta glycerolphosphate.
Refer :
Chondrogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells: Tips and Tricks
Luis A. Solchaga, Kitsie J. Penick, and Jean F. Welter
Follow these steps:
1. Chondrogenic differentiation medium: Dulbecco’s Modified Eagle’s Medium, High (4.5 g/l) Glucose (DMEM-HG, Invitrogen) supplemented with 10% ITS+ Premix Tissue Culture Supplement (Becton Dickinson), 10−7 M dexamethasone (Sigma), 1 μM ascorbate-2-phosphate (Wako, Richmond, VA), 1% sodium pyruvate (Invitrogen), and 10 ng/ml transforming growth factor-beta 1 (TGF-β1, Peprotech, Rocky Hill, NJ).
2. Dexamethasone stock solution (100×): dexamethasone (Sigma) is dissolved at 10−3 M in absolute ethanol and diluted 1:100 with DMEM-HG to obtain the 10−5M stock solution. Store at −20°C.
3. Ascorbate-2-phosphate stock solution (100×): Prepare 100 μM ascorbate-2-phosphate (Wako) in H2O. Store at −20°C.
4. Transforming Growth Factor-beta 1 (TGF-β1): Prepare a 1-μg/ml stock solution (100×) of TGF-β1 (Peprotech, Rocky Hill, NJ) in 4 mM hydrochloric acid (HCl) and 1% bovine serum albumin. Store at −80°C.
5. Polypropylene 96-well plates (Phenix, Candler, NC).
6. Electronic repeater pipette (Brand, Essex, CT).
7. Repeater pipette tips (Brand; Eppendorf, Westbury, NY).
8. Large orifice 200-μl pipette tip (Fisher).
Or , follow the method of gene induced , This reference may help you .
1. For chondrogenic differentiation it needs the cells to be cultured in a 3D compact pellet.
2. Mesenchymal stem cells isolated from different sources have different capacity to undergo chondrogenesis, hence choosing right source is important.
3. Isolation of mesenchymal stem cells should be verified by flowcytometry to ensure the purity of this hetrogen! source of stem cells. It affects their differentiation.
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