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  本文選題：骨髓間充質(zhì)干細胞 + 肝細胞生長因子�。� 參考：《廣西醫(yī)科大學》2006年碩士論文

【摘要】： 目的:探討體外誘導大鼠骨髓間充質(zhì)干細胞(bone mesenchymal stem cells , BMSCs)定向分化為肝樣細胞的可行性。方法:肝細胞生長因子(hepatocte growth factor ,HGF)體外誘導大鼠BMSCs,誘導第0d、7d、14d、21d,分別觀察細胞形態(tài)特征;將培養(yǎng)出的細胞進行堿性磷酸酶染色;免疫細胞化學法檢測甲胎蛋白(AFP)、細胞角質(zhì)蛋白18(CK18)及白蛋白(ALB)的表達;糖原染色檢測細胞糖原的表達;分別將基礎(chǔ)培養(yǎng)基培養(yǎng)21d及用HGF定向誘導21d的細胞進行透射電鏡觀察其超微結(jié)構(gòu)的變化。結(jié)果:HGF誘導組細胞從長梭形逐步轉(zhuǎn)變?yōu)閳A形,失去放射狀排列的特點;非誘導組細胞呈長梭形生長,放射狀或旋渦狀排列。培養(yǎng)至第7d的細胞進行堿性磷酸酶染色呈陽性。HGF誘導組:第7d時AFP表達增加,但此后表達逐漸減少;CK18、ALB及糖原隨誘導時間表達逐漸增強。在非誘導組:ALB、CK18、糖原在各時間點均呈陰性,AFP第0 d有少量表達,此后不再表達。定向誘導21d后的骨髓干細胞電鏡下發(fā)現(xiàn)細胞呈肝索樣排列,細胞間有大量的微絨毛,細胞內(nèi)可見到豐富的粗面內(nèi)質(zhì)網(wǎng)、線粒體、溶酶體、吞飲泡,極類似于大鼠肝細胞電鏡下的結(jié)構(gòu)。結(jié)論:BMSCs在HGF誘導下分化為肝樣細胞,肝細胞標志物表達與肝細胞發(fā)育過程吻合;其形態(tài)學上符合肝細胞的超微結(jié)構(gòu)。
[Abstract]:Aim: to investigate the feasibility of inducing rat bone mesenchymal stem cells (BMSCs) to differentiate into hepatoid cells in vitro. Methods: hepatocyte growth factor (HGF) was used to induce rat BMSCs in vitro, and to observe the morphological characteristics of BMSCs on the 7th day, 14d and 21d, respectively, and the cultured cells were stained with alkaline phosphatase (ALP). Immunocytochemistry was used to detect the expression of alpha-fetoprotein (AFP), cytokeratin 18 (CK18) and albumin (ALB), and glycogen staining was used to detect the expression of glycogen. The ultrastructural changes of the cells cultured in basic culture medium for 21 days and induced by HGF for 21 days were observed by transmission electron microscope. Results the cells of the group induced by 1% HGF gradually changed from spindle shape to round shape and lost the characteristic of radial arrangement, while the cells in non-induced group grew in long spindle shape, radial or swirl shape. The expression of AFP increased on the 7th day, but the expression of CK18, ALB and glycogen increased gradually with the induction time. In the uninduced group, no expression of glycogen was found on day 0 of AFP at all time points, but no expression was found after that. Under electron microscope of bone marrow stem cells induced by directional induction for 21 days, the cells were found to be arranged like hepatic cord, with a large number of microvilli between the cells, and abundant rough endoplasmic reticulum, mitochondria, lysosomes, and swallowing vesicles were found in the cells. Very similar to the structure of rat hepatocytes under electron microscope. Conclusion HGF induced BMSCs differentiated into hepatocyte-like cells, and the expression of hepatocyte markers coincided with the development of hepatocytes, and its morphology was in accordance with the ultrastructure of hepatocytes.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R329.2

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