Chondrogenic differentiation media (10X)
Genekam Chondrogenic Differentition Media (Sterile Product)*
Kit contents: STEM10 Genekam Chrondrogenic differentition media (10X) 10 ml (store it at -20°C)
Prepare the media while adding 10 ml of Genekam Chrondrogenic differentiation media into 90ml of DMEM containing 10 % FCS + 1% SP + 1% Glutamine or in your culturing media. Store it at 4°C. You can prepare less media, but the ratio should be 1:10.
After you have stem cells e.g. MSC. To isolate MSC cells, one can use Genekam Stem cell isolator kit. After you get 60-85% confluence during culturing of stem cells, please remove the cell using trypsin or Genekam cell remover and wash the cells to make free from trypsin while Centifuge at 1400 rpm for 5 minutes and suspend the pellet now to make the cell count using trypan blue.
Now add the cells 0.5 x 105 / ml in chrondrogenic differentiation media (containing DMEM + 10 % FCS or other serum source) in 15ml sterile tube and centrifuge it at 1200 rmp for 5 minutes. The pellet will be formed. Now keep the tube in CO2 inocubator at 5% CO2 for two weeks (donot distrub the pellet; donot tight (keep it loose!) the cap of tube in order to low the exchange of CO2) while feeding the cells 3-5 days with this media for 2 weeks.
After two weeks, you can stain with cells with Toludine blue to detect the secretion of sulfated glycosaminoglycans or stain with safranin O staining.
* only for research use.