Dear Carl,

In my hand, the easisest way to do is to make pellets with you MSC in a 15 ml conical falcon tube. Following centrifugation, simply detach the pellet from the bottom of the tube. Remove the medium and add the differentiation medium (Dulbecco’s Modified Eagle’s Medium, High (4.5 g/l) Glucose supplemented with 10-7 M dexamethasone,

50 mg/mL ascorbate, 50 mg/mL Insulin-transferrin sodium selenite, 100 mg/mL sodium pyruvate, 40 mg/mL proline, 2 mM L-glutamine, and 10 ng/mL TGF-b. After 21 days, you will have nice chondrocytes.

Best regards,

Thomas

Dear Carl,

In my hand, the easisest way to do is to make pellets with you MSC in a 15 ml conical falcon tube. Following centrifugation, simply detach the pellet from the bottom of the tube. Remove the medium and add the differentiation medium (Dulbecco’s Modified Eagle’s Medium, High (4.5 g/l) Glucose supplemented with 10-7 M dexamethasone,

50 mg/mL ascorbate, 50 mg/mL Insulin-transferrin sodium selenite, 100 mg/mL sodium pyruvate, 40 mg/mL proline, 2 mM L-glutamine, and 10 ng/mL TGF-b. After 21 days, you will have nice chondrocytes.

Best regards,

Thomas

Dear Carl, 

I have a receipt to prepare chondrogenic diff. medium for MSC: 

DMEM/F12, 10%FCS, 10-7M dexamethasone, standard Pen/Strep, set the pH by adding NaHCO3, sterilize by filtering. Add freshly 5ng/ml TGF-b3 and 6.25ug/ml insulin. (Reconstitute the twolatest according to the instructions of the manufacturer.

Alternatively you can buy this:

https://www.thermofisher.com/order/catalog/product/A1007101

I use the same protocol than Thomas except that I add tgf beta 3 instead of 1.

Hello

For chondrogenic differentiation, 2.5 × 105 MSCs were centrifuged (150 g, 5 minutes) to form high-density micromass culture pellets. The chondrogenic differentiation of these pellets was achieved for 28 days with DMEM (4.5 g/L glucose; Biochrom), ITS supplements, 100 nM dexamethasone, 0.17 mM ascorbic acid-2-phosphate, 1 mM sodium pyruvate, 0.35 mM L-proline (all Sigma-Aldrich) and 10 ng/ml transforming growth factor-β3 (TGF-β3; PeproTeck, Hamburg, Germany) 

Ref in links

Good Luck 

https://stemcellres.biomedcentral.com/articles/10.1186/scrt31

http://link.springer.com/article/10.1007/s10529-010-0293-x

http://www.promocell.com/fileadmin/promocell/PDF/Chondrogenic_Differentiation_and_Analysis_of_MSC.pdf

https://fscimage.fishersci.com/cmsassets/downloads/segment/Scientific/pdf/Cell_Culture/Protocols/Stem_Cell_Specific/chondrogenic_diffhMSCs.pdf