体外培养状态下新西兰白兔MSC的传代增殖特性  

Observations on the Proliferation in Vitro of Subcultured MSCin New Zealand White Rabbits

作　　者：刘丹平[1] 栗刚[1] 张德志[1] 关振家[1] 胡蕴玉[2] 郭韬[1] 张男[1] 

机构地区：[1]锦州医学院附属第一医院骨科,辽宁锦州121000 [2]西安第四军医大学西京医院全军骨科研究所,陕西西安710000

出　　处：《锦州医学院学报》2003年第2期1-4,共4页Journal of Jinzhou Medical College

摘　　要：目的　观察体外状态下新西兰白兔骨髓基质细胞 (MSC)的传代增殖特性 ,评价其体外生存能力。方法抽取新西兰白兔股骨骨髓行 1 2天原代培养后再行传代培养 8代 ,用细胞计数法测定传第 1代、第 4代、第 8代MSC的增殖情况 ,绘制生长曲线。本研究还检测了传 1代的MSC的分裂指数和细胞贴壁率。结果　传 1代MSC的生长曲线与传 4代、传 8代MSC的生长曲线基本一致 ,其生长曲线显示在接种后的第 2个～第 5个 2 4h (共 4天 )细胞增殖是呈对数生长状态 ,此时的群体倍增时间是 30 87h。培养第 7天以后细胞基本停止生长。细胞的分裂指数在传代的第 2个、第 3个 2 4h增长最快 ,第 4个 2 4h达到高峰 (2 3 8% ) ,在第 9个 2 4h细胞已处于汇合状态 ,基本停止分裂。传 1代MSC细胞贴壁率在传代后第 2h、第 8h、第 1 0h分别为 2 8 5 %、 85 0 %、92 5 % ,此后贴壁率上升趋于停止 ,表明细胞贴壁主要发生在传代后 1 0h以内。结论　新西兰白兔MSC是一种在体外状态下生长旺盛 ,生存能力较强 。Objective To investigate the proliferation in vitro of subcultured marrow stromal cells(MSC)in New Zealand white rabbits for evaluating their viability Methods The femoral marrow was aspirated for 12 day ' s primary culture,and then subculture was done with conventional techniques employing 0 25%trypsin and then 8 successive passages of MSC were subcultured in our present study.The proliferations of the lst,the 4th and the 8th of subcultured MSC were determined respectively using cell counting done in a haemocytometer,and the proliferation curves(PC)of the lst MSC passage were plotted.The mitotic index(MI)and the seeding efficiency(SE)of the lst passage MSC were also determined.Results The PC of the lst passage MSC was basicaly consistent with that of the 4th and the 8th passages.The PC revealed that the MSC inoculated in the 2nd～the 5th 24 hours(4 days)were in log proliferation phase that their population doubling time(PDT)was 30 87 hours.The proliferation ceased after 7 days inoculation.The MI went up more rapidly in the 2nd and the 3rd 24 hours and reached the peak at the 4th 24 hour(23 8%).After the cells were in entire confluence at the 9th 24 hours,the Ml reduced close to zero.The SEs of the MSC were 28 5%,85 0%and 92 5% after inoculated for 2 hours,8 hours and 10 hours respectively.The SE rise tended to stop after subcultured for the 10th hour.This demonstrastes that the majority of the cells attachment acurred within 10 hours after inoculation. Conclusions As one kind of osteogenic potential cells,MSC of New Zealand white rabbit are in vitro viable and eugonic.

关 键 词：骨髓基质细胞 传代增殖特性 体外培养状态 新西兰白兔 体外生存能力 原代培养 

分 类 号：R331.14[医药卫生—人体生理学] R329－33[医药卫生—基础医学]