p(3HB-co-3HH)对哺乳类动物骨髓干细胞生物学行为的影响

The effects of p(3HB-co-3HH) on biobehavior of mammiferous BMSCs

目的 :通过观察 3 羟基丁酸、3 羟基己酸嵌段共聚 [poly(3 hydroxybutyrate co 3 hydroxyhexanoate) ,p(3HB co 3HH) ]对犬骨髓干细胞生物学行为的影响 ,探索 p(3HB co 3HH)作为组织工程支架材料在哺乳类动物体内应用的可行性。方法 :应用p(3HB co 3HH)在DMEM培养液中的浸出液 (0 1g/ml)培养犬骨髓干细胞 ,绘制细胞生长曲线 ,MTT测定及用FCM对细胞周期时相进行分析 ,并在 p(3HB co 3HH)薄膜上培养骨髓干细胞 ,观察细胞形态。结果 :在细胞培养的第 2～ 5d ,细胞增殖明显 ;MTT测定相对增殖率 10 2 % ,细胞毒性 0级 ;FCM检测结果显示p(3HB co 3HH)浸出液培养组增殖期细胞为 2 2 18% ,对照组为 16 12 %。骨髓干细胞在 p(3HB co 3HH)薄膜上培养 48h后观察呈簇状粘附生长。结论 :p(3HB co 3HH)浸出液对犬骨髓干细胞无毒性 ,细胞在 p(3HB co 3HH)表面生长良好 。

Objective:This study observed the effects of poly[(3 hydroxybutyrate co 3 hydroxyhexanoate),p(3HB co 3HH)]on biobehavior of mammiferous bone marrow stem cells(BMSCs) and the feasibility of p(3HB co 3HH) as a new type of biodegradable materials in tissue engineering Methods:The BMSCs of canine were cultured in the lixivium of p(3HB co 3HH) in DMEM containing 15% fetal bovine serum Cell growth curve was protracted according to the sum of cells every day The toxicity of the lixivium to BMSCs was mensurated using MTT test The stages of cell cycle were analyzed with flow cytometry(FCM) The configuration of BMSCs inoculated on the film of p(3HB co 3HH) was observed under microscope Results: The cell growth curve showed that the sum of the cells cultured with the lixivium was higher than cultured without the lixivium from 2 to 5 days In the presence of the lixivium , the relative growth rate(RGR) of cultured BMSCs was 102% at 96 hours(t test, P >0 05) ,and the toxicity grade of the lixivium to BMSCs was 0 scale The results of FCM indicated that BMSCs in proliferating phase was 22 18% in cultured with the lixivium and it was 16 12% in control group On the film of p(3HB co 3HH),the appearance of BMSCs was fibroblastoid and clustered Conclusions:The lixivium of p(3HB co 3HH)was innocuous to mammiferous BMSCs and prompted BMSCs to proliferate In the future, p(3HB co 3HH) may be applied in tissue engineering