本文選題：骨髓 + 間充質(zhì)干細(xì)胞　； 參考：《華中科技大學(xué)》2007年碩士論文

【摘要】： 目的:研究大鼠骨髓間充質(zhì)干細(xì)胞(rMSCs)的體外分離培養(yǎng)、擴(kuò)增及其向視網(wǎng)膜神經(jīng)細(xì)胞分化的潛能。 方法:取4-6周SD大鼠雙側(cè)股骨和脛骨骨髓用培養(yǎng)基懸浮,接種于FN包被的六孔板中,純化并擴(kuò)增rMSCs,顯微鏡下觀察細(xì)胞生長(zhǎng)狀態(tài)及形態(tài)學(xué)變化,并用流式細(xì)胞儀檢測(cè)其表型;同時(shí)取生后1-3天SD乳鼠視網(wǎng)膜神經(jīng)上皮層培養(yǎng),定期取上清夜用作配制分化液。通過(guò)兩步法不同的誘導(dǎo)劑誘導(dǎo)第8-10代rMSCs分化,顯微鏡下觀察分化細(xì)胞的形態(tài)學(xué)變化,免疫熒光細(xì)胞化學(xué)檢測(cè)分化過(guò)程中神經(jīng)干細(xì)胞以及視網(wǎng)膜神經(jīng)細(xì)胞部分特異性標(biāo)記蛋白的表達(dá)。 結(jié)果:rMSCs貼壁生長(zhǎng),表現(xiàn)為成纖維細(xì)胞樣,呈梭形、多角形等細(xì)胞形態(tài),達(dá)到融合時(shí)呈長(zhǎng)梭形緊密排列,形態(tài)均一,成旋渦樣生長(zhǎng),20代以后細(xì)胞生長(zhǎng)逐漸衰退、出現(xiàn)分化傾向、形態(tài)異質(zhì);表型分析結(jié)果顯示rMSCs表達(dá)CD44,CD90不表達(dá)CD34,CD45;第一步誘導(dǎo)分化后,細(xì)胞形態(tài)改變并逐漸交織成網(wǎng)狀,最后形成球形,不斷增大,呈現(xiàn)神經(jīng)干細(xì)胞樣生長(zhǎng)特性,并高表達(dá)其標(biāo)志nestin,第二步誘導(dǎo)后細(xì)胞呈雙極、多極或錐形,表達(dá)NeuN、GFAP,部分表達(dá)視網(wǎng)膜神經(jīng)節(jié)細(xì)胞標(biāo)志Thy1.1。 結(jié)論:rMSCs可以在體外成功分離培養(yǎng)、擴(kuò)增純化,具有向神經(jīng)干細(xì)胞分化的潛能,并可進(jìn)一步分化出視網(wǎng)膜神經(jīng)樣細(xì)胞,可用作臨床藥物實(shí)驗(yàn)研究,并有望成為移植治療視網(wǎng)膜神經(jīng)細(xì)胞病變的工具細(xì)胞。
[Abstract]:Aim: to study the potential of rat bone marrow mesenchymal stem cells (MSCs) to differentiate into retinal nerve cells in vitro. Methods: bone marrow of bilateral femur and tibia of 4-6 weeks Sprague-Dawley rats were suspended in culture medium and inoculated into FN-coated six-hole plate. RMSCs were purified and amplified. The growth state and morphological changes of the cells were observed under microscope and the phenotypes were detected by flow cytometry. At the same time, 1-3 days after birth SD rats retinal nerve epithelium culture, and regular supernatant night for preparation of differentiation fluid. The differentiation of 8-10 passage rMSCs was induced by different inducers of two-step method. The morphological changes of differentiated cells were observed under microscope. The expression of partial specific labeled proteins in neural stem cells and retinal nerve cells during differentiation was detected by immunofluorescence cytochemistry. Results the adherent cells of 10% rMSCs appeared as fibroblasts, fusiform, polygonal, and so on. When they reached fusion, they were arranged closely in long spindle shape and uniform in shape. After 20 generations of vortex like growth, the cell growth gradually declined, and the differentiation tendency appeared. Phenotypic analysis showed that the expression of CD44T CD90 was not expressed in rMSCs. After the first step of differentiation, the cell morphology changed and gradually intertwined into reticular shape. Finally, the cells became globular and gradually enlarged, showing the characteristics of neural stem cell like growth. After the second step, the cells showed bipolar, multipolar or conical expression, and some of them expressed NeuNa-GFAPs, and some of them expressed Thy1.1, a marker of retinal ganglion cells. Conclusion\%\ It is expected to be a tool cell for the transplantation of retinal neuropathy.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 金澎,徐如祥,姜曉丹,杜謀選,蔡穎謙,鄒雨汐;骨髓源性神經(jīng)干細(xì)胞體外培養(yǎng)體系的安全性評(píng)價(jià)[J];中華神經(jīng)醫(yī)學(xué)雜志;2005年01期

2 楊智寬,葛堅(jiān),郭彥,王智崇,劉海泉;SD大鼠視網(wǎng)膜神經(jīng)細(xì)胞培養(yǎng)上清液對(duì)胚胎干細(xì)胞體外分化的誘導(dǎo)作用[J];中華眼底病雜志;2002年02期

，

本文編號(hào)：1949295