骨折预处理对兔骨髓来源间充质干细胞的影响

目的探讨骨折预处理对兔骨髓来源间充质干细胞（MSC）的影响。方法将10只兔按随机数字表法分为2组,每组5只。骨折预处理组截断兔股骨骨干后采用克氏针内固定,7 d后从另一侧股骨无菌抽取骨髓1～2 mL;对照组未进行任何处理。采用全骨髓贴壁法对2组兔进行骨髓来源的间充质干细胞的原代培养、酶消化法进行人工纯化,通过形态学观察、细胞周期测定及MTT实验,研究骨折预处理对MSC的影响。结果骨折预处理后可较快地获得纯化的MSC,且MSC的增殖能力在P2、P3代时均高于对照组（均P〈0.05）;随着时间的延长,P4代时2组MSC增殖能力比较差异无统计学意义（P〉0.05）。对P4代进行细胞周期测定,骨折预处理组MSC中G0/G1期细胞为70.65%,对照组MSC中G0/G1期细胞为72.40%,2组比较差异无统计学意义（P〉0.05）。结论骨折预处理全骨髓贴壁法可在更短的时间内成功地培养出稳定的MSC,并且该细胞具有更强的增殖能力。

Influence of Pretreatment of Bone Fracture on Bone Marrow-derived Mesenchymal Stem Cells in Rabbits

Objective To investigate the influence of the pretreatment of bone fracture on bone marrow-derived mesenchymal stem cells（MSCs）in rabbits.Methods Ten rabbits were randomly divided into two groups,with 5 rabbits in each group.In pretreatment group,Kirschner wire fixation was performed after femoral shaft fracture and 1-2 mL of bone marrow was aseptically aspirated from the other side of the femur bone after 7 days.Control group received no treatment.Bone marrow-derived MSCs were cultured and purified by whole bone marrow adherent method and enzyme digest method.The effect of the pretreatment of bone fracture on bone marrow-derived MSCs was determined by morphological observation,cell cycle analysis and MTT experiment.Results Compared with control group,MSCs were quickly purified and the proliferation of cells at passage 2 and 3 significantly increased in pretreatment group（P〈0.05）.However,no obvious differences in cell proliferation at passage 4 were found between the two groups（P〉0.05）.Moreover,there were no significant differences in the number of G0/G1 phase cells between pretreatment group and control group（70.65% vs 72.40%,P〉0.05）.Conclusion Bone marrow-derived MSCs can be quickly purified and cell proliferation can be significantly increased by the pretreatment of bone fracture.