In vitro culture and cell cycle detection of the adult degenerated nucleus pulposus / 中国矫形外科杂志

ABSTRACT

[Objective]To establish an in vitro two-dimensional culture model of degenerated nucleus pulposus and detect samples' cell cycle by flow cytometry to study why nucleus pulposus cells don't grow well after passage. [Method]Nucleus pulposus tissues taken from protruded discs of 24 patients were treated by Trypsin and collagenase after surgical procedures,and then the cells were cultured in DMEM/F12 medium.Cell morphology was observed by an invert microscope and cell cycle of the primary and P2 cells were detected by flow cytometry after proliferation in monolayer culture.[Result]1.Primary culture cells of nucleus pulposus grew well in the medium,and 90% cells adhere to layer after about 7d.2.The rate of apoptosis of NP primary(38.10?11.7)%,P2(44.74?17.6)%.The rate of S period primary(7.88?2.1%),P2(2.76?0.7)%.[Conclusion]When going down to posterity the cells' apoptosis rate grows while S period cells decrease.