本文選題：睪丸支持細(xì)胞　切入點(diǎn)：骨髓基質(zhì)細(xì)胞　出處：《廣西醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 目的: 分離培養(yǎng)睪丸支持細(xì)胞、骨髓基質(zhì)細(xì)胞、胚胎成纖維細(xì)胞和胰腺成纖維細(xì)胞,優(yōu)化培養(yǎng)條件,建立飼養(yǎng)細(xì)胞系,為干細(xì)胞體外共培養(yǎng)體系的建立提空充足的飼養(yǎng)細(xì)胞。 方法: 1.使用膠原酶消化法和低滲處理分離睪丸支持細(xì)胞(Sertoli cell,SC);全骨髓離心后貼壁培養(yǎng)分離骨髓基質(zhì)細(xì)胞(Bone marrow stromalcell,BMSC),多種培養(yǎng)液對比培養(yǎng);組織消化法分離培養(yǎng)小鼠胚胎成纖維細(xì)胞(Mouse embryonic fibroblast,MEF)和胰腺成纖維細(xì)胞(PancreaticFibroblast,PF)。 2.MTF法測定第三代胚胎成纖維細(xì)胞和同代胰腺成纖維細(xì)胞的生長活力,使用DMSO低溫保存細(xì)胞;分別取第三代的胚胎成纖維細(xì)胞和胰腺成纖維細(xì)胞,加入10μg/ml的絲裂霉素C制備飼養(yǎng)層。 結(jié)果: 低滲處理可以獲得富集度較高的SC;含15%FBS的DMEM(L)培養(yǎng)液更適合BMSC體外培養(yǎng);經(jīng)組織塊貼壁培養(yǎng)法從胰腺結(jié)締組織中分離獲得成纖維樣細(xì)胞,此類細(xì)胞具有成纖維細(xì)胞生長活性;使用DMSO作為細(xì)胞保護(hù)劑凍存的MEF和PF,復(fù)蘇后可繼續(xù)傳代培養(yǎng);制備了MEF和PF飼養(yǎng)層。 結(jié)論: 1.低滲處理可以有效地提高睪丸支持細(xì)胞的富集度; 2.含15%FBS的DMEM(L)培養(yǎng)液更適合BMSC體外培養(yǎng); 3.從胰腺結(jié)締組織中分離PF,此細(xì)胞和MEF具有相似生物學(xué)活性; 4.初步建立了小鼠胚胎成纖維細(xì)胞系和胰腺成纖維細(xì)胞系。 5.制備了MEF和PF兩種飼養(yǎng)層。
[Abstract]:Objective:. Sertoli cells, bone marrow stromal cells, embryonic fibroblasts and pancreatic fibroblasts were isolated and cultured. Methods:. 1. Sertoli cells were isolated by collagenase digestion and hypoosmotic treatment, bone marrow stromal cells were isolated by adherent culture after whole bone marrow centrifugation. Mouse embryonic fibroblast (MEF) and pancreatic fibroblast (Pancreatic fibroblast) were isolated and cultured by tissue digestion. 2.MTF method was used to determine the growth activity of the third generation of embryonic fibroblasts and the pancreatic fibroblasts of the same generation. The cells were cryopreserved by DMSO, and the third generation of embryonic fibroblasts and pancreatic fibroblasts were obtained, respectively. The feeder layer was prepared by adding 10 渭 g/ml mitomycin C. Results:. Hypotonic treatment could obtain high concentration of SCS; the culture medium containing 15s was more suitable for BMSC culture in vitro; fibroblasts were isolated from pancreatic connective tissue by tissue mass adherent culture, and fibroblast-like cells were obtained from pancreatic connective tissue by tissue mass adherent culture. The cells had fibroblast growth activity, MEF and PFL, which were frozen by DMSO as cell protectant, could be further cultured after resuscitation. MEF and PF feeder layers were prepared. Conclusion:. 1. Hypotonic treatment can effectively increase the concentration of Sertoli cells in testis. 2. The culture medium containing 15s was more suitable for BMSC culture in vitro. 3.PFs isolated from connective tissue of pancreas showed similar biological activity to MEF. 4. Mouse embryonic fibroblasts and pancreatic fibroblasts were established. 5. Two feeding layers, MEF and PF, were prepared.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R329

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 劉海超;張巖;于泊洋;吳應(yīng)積;;制備精原干細(xì)胞滋養(yǎng)層細(xì)胞條件的優(yōu)化[J];現(xiàn)代生物醫(yī)學(xué)進(jìn)展;2012年04期

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