發(fā)布時間：2018-03-24 05:34

  本文選題：軟骨細胞　切入點：軟骨單位　出處：《山西醫(yī)科大學》2013年碩士論文

【摘要】：目的：利用兔關節(jié)軟骨細胞與軟骨單位建立混合共培養(yǎng)體系，明確體外軟骨細胞與軟骨單位共培養(yǎng)構(gòu)建軟骨組織工程種子細胞的最佳比值。 方法：將兔膝關節(jié)軟骨分別用酶消化為軟骨細胞與軟骨單位后，按一定比例混合配制，分為五組，A組:單純軟骨細胞組；B組:比值為2:1的軟骨細胞/軟骨單位組；C組:比值為1:1的軟骨細胞/軟骨單位組；D組:比值為1:2的軟骨細胞/軟骨單位組；E:單純軟骨單位組。以2×105個細胞/孔接種于放有蓋玻片的六孔板中，使細胞單層貼壁培養(yǎng)爬片，待細胞生長6日爬滿后，進行指標檢測： HE、番紅O染色觀察細胞組織形態(tài)學的變化，RT-PCR檢測Agg、II型膠原和I型膠原的基因表達水平。Elisa檢測上清液GAG、II型膠原的含量。 結(jié)果：共培養(yǎng)組細胞形態(tài)正常。通過RT-PCR檢測Agg、II型膠原和I型膠原的基因表達，Elisa檢測上清液GAG、II型膠原的含量表明：采用共培養(yǎng)方法的軟骨細胞與軟骨單位的混合組中細胞代謝的表達要明顯高于單純軟骨細胞組，尤其在軟骨細胞/軟骨單位共培養(yǎng)組（1:1），糖胺多糖與II型膠原在基因的表達和代謝情況均為最高。 結(jié)論：在軟骨組織工程移植種子細胞時，，軟骨細胞與軟骨單位為1∶1的比例為最佳選擇，適用于軟骨組織工程。
[Abstract]:Aim: to establish a mixed co-culture system of rabbit articular chondrocytes and chondrocytes to determine the optimal ratio of chondrocytes co-cultured with chondrocytes to chondrocyte seed cells for cartilage tissue engineering. Methods: the articular cartilage of rabbit knee joint was digested into chondrocytes and chondrocytes by enzyme, then mixed into chondrocytes and chondrocytes in a certain proportion. Divided into five groups: chondrocyte group B: chondrocyte / chondrocyte unit group C group with 2:1; chondrocyte / chondrocyte unit group D group with 1:1; chondrocyte / chondrocyte / chondrounit group with 1:2 ratio: single chondrocyte / chondrocyte unit group. Pure chondrocyte unit group. Inoculated with 2 脳 105 cells / hole in a six-hole plate with a cover glass, Make the cell monolayer adhere to the wall of the climbing piece, after 6 days of cell growth full, The changes of histomorphology were observed by HEand Phangnon O staining. The gene expression level of type II and type I collagen was detected by RT-PCR. Elisa was used to detect the content of type II collagen in supernatant. Results: the morphology of the co-cultured cells was normal. The gene expression of type II collagen and type I collagen was detected by RT-PCR. The content of type II collagen in supernatant was determined by Elisa. The results showed that the chondrocytes and chondrocytes were mixed with chondrocytes by co-culture method. The expression of cell metabolism in the chondrocyte group was significantly higher than that in the chondrocyte group. Especially in chondrocyte / chondrocyte coculture group (1: 1), glycosaminoglycan and type II collagen had the highest gene expression and metabolism. Conclusion: the ratio of chondrocytes to chondrocytes is 1:1, which is suitable for cartilage tissue engineering.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R329

【參考文獻】

相關期刊論文 前4條

1 林薇薇;陳金武;石利強;孟慶江;王美青;史慶輝;;漸進性咬合紊亂對髁突軟骨I型膠原表達的影響[J];臨床口腔醫(yī)學雜志;2008年12期

2 段王平;孫振偉;衛(wèi)小春;;關節(jié)軟骨中最基本的解剖功能結(jié)構(gòu):軟骨單位(英文)[J];中國組織工程研究與臨床康復;2010年24期

3 段王平;孫振偉;李琦;任立新;衛(wèi)小春;;兔膝關節(jié)軟骨單位體外酶解法消化 分離的實驗研究[J];中國藥物與臨床;2010年08期

4 黃永波,衛(wèi)小春,翁習生,尹昆,李鵬翠,陸向東;海藻酸鈉-成年軟骨細胞培養(yǎng)移植修復成年兔關節(jié)軟骨缺損的研究[J];中華實驗外科雜志;2004年08期

相關博士學位論文 前1條

1 段王平;不同年齡兔膝關節(jié)軟骨單位生物學特性分析[D];山西醫(yī)科大學;2011年

本文編號：1656931