非接触共培养条件下山羊BMSCs向纤维环样细胞诱导分化的研究?

发布时间：2018-08-03 06:47

【摘要】：[目的]建立山羊骨髓源性间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)和纤维环细胞(annulus fibrosus cells,AFCs)非接触共培养体系,探讨BMSCs向纤维环样细胞诱导分化潜能,为组织工程修复纤维环缺损研究提供种子细胞。[方法]复苏第3代AFCs和BMSCs,观察细胞形态并绘制生长曲线。采用Transwell系统共培养AFCs和BMSCs,7 d后,检测BMSCs向纤维环样细胞分化情况。通过甲苯胺蓝染色检测聚集蛋白聚糖的表达,通过免疫细胞化学法检测Ⅰ型胶原蛋白的表达,realtime PCR检测I型胶原、聚集蛋白聚糖和Sox-9 m RNA表达。[结果]非接触共培养7 d后,BMSCs形态转变为纤维环细胞样。非接触共培养组中BMSCs的聚集蛋白聚糖与I型胶原染色呈阳性,与AFCs对照组结果一致,BMSCs对照组结果呈阴性。与AFCs对照组相比,共培养组中I型胶原、聚集蛋白聚糖及Sox-9的m RNA表达量分别为0.86、1.13和0.91(P0.05),而BMSCs对照组中未检测到其表达。[结论]建立稳定的山羊AFCs和BMSCs非接触共培养体系,共培养7 d后,BMSCs具有AFCs特性,表明共培养条件促进BMSCs诱导分化为纤维环样细胞。
[Abstract]:[objective] to establish a non-contact co-culture system between goat bone marrow-derived mesenchymal stem cells (bone marrow mesenchymal stem cells) and fibroannular cells (annulus fibrosus cells), and to explore the differentiation potential of BMSCs into fibrocyte-like cells. To provide seed cells for tissue engineering repair of fibrous ring defect. [methods] the third generation of AFCs and BMSCs were resuscitated to observe the morphology and draw the growth curve. After 7 days of co-culture of AFCs and BMSCs with Transwell system, the differentiation of BMSCs into fibroid cells was detected. The expression of type I collagen, aggregate proteoglycan and Sox-9 m RNA were detected by toluidine blue staining, and the expression of type 鈪，

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