纤维蛋白胶对脂肪干细胞增殖和存活的影响

Effects of fibrin glue on the proliferation and viability of adipose tissue-derived stem cells

背景:尽管有很多生物支架材料被用于携带种子细胞,但纤维蛋白胶与脂肪干细胞二者在体外的共培养研究及种子细胞在支架材料中的增殖存活评价没有统一的标准。目的:探讨纤维蛋白胶对体外培养的大鼠脂肪干细胞的增殖和存活性,为可注射组织工程化心肌治疗心肌梗死提供实验依据。方法:分离培养大鼠脂肪干细胞,通过流式细胞术检测其免疫表型,将脂肪干细胞与纤维蛋白胶共培养,用噻唑蓝比色法及LIVE/DEAD荧光双染色检测脂肪干细胞在纤维蛋白胶中的增殖和存活性。结果与结论:大鼠脂肪干细胞表型特征为CD90、CD105阳性,CD34、CD45阴性,噻唑蓝比色结果显示,纤维蛋白胶中的脂肪干细胞增殖活性高于常规培养组(P<0.05),LIVE/DEAD荧光双染色显示脂肪干细胞在纤维蛋白胶中生长良好,24,72h的细胞存活率均在90%以上,死亡率小于7%。结果提示,大鼠脂肪干细胞在纤维蛋白胶中增殖和存活良好,可用纤维蛋白支架携带脂肪干细胞用于可注射心肌组织工程研究。

BACKGROUND：Many biological scaffolds are used in carrying seed cells,however,there is no criterion about the co-culture of fibrin glue with adipose tissue-derived stem cells and the evaluation of the proliferation and viability of seed cells on the scaffolds.OBJECTIVE：To investigate the effects of fibrin glue on the proliferation and viability of rat adipose tissue-derived stem cells cultured in vitro,and to provide the experimental evidence for injectable tissue engineered myocardial treatment for myocardial infarction.METHODS：Adipose tissue-derived stem cells were isolated from rats and then cultured.Their immunophenotype was identified by flow cytometry.Adipose tissue-derived stem cells were co-cultured with fibrin glue.The proliferation of adipose tissue-derived stem cells were tested by MTT assay and cell viability was assessed with LIVE/DEAD fluorescence double stain.RESULTS AND CONCLUSION：The adipose tissue-derived stem cells of rats were positive for CD90 and CD105,but negative for CD34 and CD45.MTT assay showed that adipose tissue-derived stem cells proliferated faster in fibrin glue group than the control group（P 0.05）;LIVE/DEAD fluorescence double stain showed that adipose tissue-derived stem cells grew well in fibrin glue.The viability of adipose tissue-derived stem cells maintained 90% and the mortality was 7% at 24 hours and 72 hours.Adipose tissue-derived stem cells well proliferate and survive in fibrin glue,fibrin scaffold may work as a carrier in deliverying adipose tissue-derived stem cells for injectable myocardial tissue engineering study.