Xiaobi Spirit Vitro Induced Rat Bone Marrow Mesenchymal Stem Cells Into Chondrocytes Differentiation Observed

Objective: Once articular cartilage is destroyed, cartilage cell can notregenerate because there is no sufficient blood supply or undifferentiatedcell. cartilage tissue has its own distinct physiological features:no bloodvessel, no nerve, no lymph draining, in articular cavity, it lives onlyon synovial fluid. Its metabolism style is mainly no-oxygen zymohydrolysis,and this decides its regeneration capability is enormously restricted, soclinically it is difficult to cure tone article。To follow our countrypopulation aging, gerontism hypertrophic arthritis are increasingly, weare worth for intimate show solicitude and explore to solve this problem.This experiment approach rat-MSCs which are fractional cultivation,generation in vitro, and in Chinese crude drug circumstance to derivationrat-MSCs chondroblast differentiated, to explore the front view ofcartilage coloboma architecture engineering recovery by Chinese crudedrug。Method: To take rat ambi-femoral bone marrow, 70%1ml Percoll FL 1200r/mincentrifuge to gain MSCs. Inoculating in 75ml culture flask which contain15%FBS Lo CHO DMEM。Put in 37℃,5%CO₂ and saturated humidity incubatorto conventional culture。After 24 hour first time to change FL, removalnulli-adherent cell, afterward per demi-2days change FL。cell growth to;80%-90% coalesce passage adoption generation 3～5 cells to divide groups。Result: (1)The primary cultured MSCs would get 80～90% confluence within2 weeks generally. The offspring cells grew more rapidly and would get 80—90% confluence within 5 to 7 days. (2) The third passage cells fromdifferent age rats oth regenerated. There would be incubation time for 1 —2 days, then the ells would exponentially grow for 3—4 days. At lastthese cells' 'quantity would reach a platform. The doubling generatingtimes were both short and only 3—4 days. There was no discrepancy in thegmwth pattern of the third passage MSCs between different age rats.Conclusion: High purity MSCs could be obtained by density with Percollfluid. The MSCs accrementited active in vitro, gradient centrifugation。...