【摘要】：[目的]建立山羊骨髓源性間充質(zhì)干細(xì)胞(bone marrow mesenchymal stem cells,BMSCs)和纖維環(huán)細(xì)胞(annulus fibrosus cells,AFCs)非接觸共培養(yǎng)體系,探討B(tài)MSCs向纖維環(huán)樣細(xì)胞誘導(dǎo)分化潛能,為組織工程修復(fù)纖維環(huán)缺損研究提供種子細(xì)胞。[方法]復(fù)蘇第3代AFCs和BMSCs,觀察細(xì)胞形態(tài)并繪制生長曲線。采用Transwell系統(tǒng)共培養(yǎng)AFCs和BMSCs,7 d后,檢測BMSCs向纖維環(huán)樣細(xì)胞分化情況。通過甲苯胺藍(lán)染色檢測聚集蛋白聚糖的表達(dá),通過免疫細(xì)胞化學(xué)法檢測Ⅰ型膠原蛋白的表達(dá),realtime PCR檢測I型膠原、聚集蛋白聚糖和Sox-9 m RNA表達(dá)。[結(jié)果]非接觸共培養(yǎng)7 d后,BMSCs形態(tài)轉(zhuǎn)變?yōu)槔w維環(huán)細(xì)胞樣。非接觸共培養(yǎng)組中BMSCs的聚集蛋白聚糖與I型膠原染色呈陽性,與AFCs對照組結(jié)果一致,BMSCs對照組結(jié)果呈陰性。與AFCs對照組相比,共培養(yǎng)組中I型膠原、聚集蛋白聚糖及Sox-9的m RNA表達(dá)量分別為0.86、1.13和0.91(P0.05),而BMSCs對照組中未檢測到其表達(dá)。[結(jié)論]建立穩(wěn)定的山羊AFCs和BMSCs非接觸共培養(yǎng)體系,共培養(yǎng)7 d后,BMSCs具有AFCs特性,表明共培養(yǎng)條件促進(jìn)BMSCs誘導(dǎo)分化為纖維環(huán)樣細(xì)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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