Induction Osteogenic differentiation medium /Osteogenic differentiation medium

Mesenchymal stem cell osteogenic differentiation kit is a self-developed low-serum medium for human mesenchymal stem cells to induce osteogenic differentiation. Cells cultured in this medium can be used for inducing osteogenic differentiation of mesenchymal stem cells from bone marrow, umbilical cord, fat and other tissues. 

Medium formula: Basic medium: 500ML, cell growth factor(MSGCS): 5ml(P2002), osteoblast induction factor: 5ml(P1303), fetal bovine serum: 25ml (SD0200), G/A: 5ml(SD0038) 

l The induction and differentiation procedure is simple and convenient. 

l is efficient in inducing osteoblasts. 

Bone marrow mesenchymal stem cell induction (Day12) 

Osteogenesis induction step:

One: Use cell cell coating solution to coat cell culture plate, and inoculate subcultured or resuscitated frozen mesenchymal stem cells onto 6-well plate with density of 2 × 105 -3 × 105 cells/cm2 or 2 × 105 -3 × 105 cells/well, cultured in a 37 ℃, 5% CO2 incubator. When the cell confluence reaches 80% - 90%, carefully suck out the mesenchymal stem cell culture medium in the well, and add 2mL of mesenchymal stem cell osteogenic differentiation medium into the 6-well plate, which is recorded as the 0 th day;

Note: Osteoblastic differentiation is easy to cause cells to roll up. During osteoblastic differentiation, the cell culture plate should be coated with coating solution, and polylysine solution (product number: SD0042) can be used for cell coating.

Two: Replace fresh osteoblastic differentiation medium of mesenchymal stem cells every 1-2 days;

Note: In order to prevent osteoblasts from falling off, it is suggested that after a large number of calcium nodules appear in the process of osteogenesis, the form of fluid change should be changed to one and a half times a day.

Three: During 14-21 days of induction, the cells were stained according to their morphological changes and growth.

Four :After the induction of osteogenic differentiation, suck the osteogenic differentiation medium from the 6-well plate, and rinse it with PBS for 1-2 times. Add 1 mL of cell fixation solution into each well and fix for 10~30 minutes;

Five: Suck off the cell fixative and wash it with PBS twice. Add 2 mL of osteoblastic test dye solution to each well and dye for 3-5 min;

Six :Suck off the staining solution for bone formation test, and wash it with PBS for 2-3 times;

Seven:Place the culture plate under the microscope to observe the staining effect of osteogenesis

Statement: The product is for scientific research only.

1. In vitro biocompatibility and stem cell regenerative assessment of hollow hydroxyapatite spheres deposited wollastonite/Ca2P6O17/TCP/ doped-wollastonite scaffolds ,https://www.sciencedirect.com/science/article/pii/S0272884224030219