大鼠骨髓和脂肪間充質(zhì)干細(xì)胞增殖及神經(jīng)分化能力對(duì)比研究
發(fā)布時(shí)間:2017-12-30 23:38
本文關(guān)鍵詞:大鼠骨髓和脂肪間充質(zhì)干細(xì)胞增殖及神經(jīng)分化能力對(duì)比研究 出處:《新疆醫(yī)科大學(xué)》2014年碩士論文 論文類型:學(xué)位論文
更多相關(guān)文章: 骨髓間充質(zhì)干細(xì)胞 脂肪間充質(zhì)干細(xì)胞 細(xì)胞分化 神經(jīng)球 神經(jīng)元
【摘要】:目的:比較骨髓間充質(zhì)干細(xì)胞和脂肪間充質(zhì)干細(xì)胞的增殖、向神經(jīng)干細(xì)胞誘導(dǎo)及分化為神經(jīng)元等終末細(xì)胞的能力。方法:取同一SD大鼠的骨髓間充質(zhì)干細(xì)胞(Bone marrow mesenchymal stem cells,BMSCs)和脂肪間充質(zhì)干細(xì)胞(Adipose-derived strom cells,ADSCs)體外分離培養(yǎng),對(duì)其形態(tài)學(xué)、增值速度進(jìn)行比較。然后將兩種間充質(zhì)干細(xì)胞誘導(dǎo)為神經(jīng)球并分化為神經(jīng)元,用CCK-8檢測(cè)神經(jīng)球的增殖速度,并計(jì)算神經(jīng)球成球率,通過免疫熒光技術(shù)比較兩種間充質(zhì)干細(xì)胞誘導(dǎo)及分化為終末神經(jīng)細(xì)胞的差異,利用ELISA檢測(cè)BDNF和NT-3神經(jīng)營養(yǎng)因子的表達(dá)情況。結(jié)果:兩種間充質(zhì)干細(xì)胞形態(tài)上稍有不同,兩種細(xì)胞均可通過傳代達(dá)到細(xì)胞純化,ADSCs比BMSCs更快達(dá)到誘導(dǎo)所需的細(xì)胞純度。生長曲線上可見ADSCs比BMSCs增值更快。BMSCs和ADSCs均易于用分步法誘導(dǎo)為神經(jīng)球及分化為終末神經(jīng)細(xì)胞,ADSCs和BMSCs成神經(jīng)球率分別為21.734±3.46%和15.62±2.37%,ADSCs明顯高于BMSCs(P0.05),且成神經(jīng)球速度快于BMSCs。免疫熒光染色結(jié)果顯示ADSCs β-tubulinⅢ陽性率(22.18±8.91%)明顯高于BMSCs(17.98±11.43%),(P0.05)。ELISA檢測(cè)結(jié)果說明誘導(dǎo)前后ADSCs比BMSCs分泌更多的BDNF和NT-3,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:ADSCs較BMSCs取材方便,增殖更快,神經(jīng)分化率更高,并且能夠分泌更多的神經(jīng)營養(yǎng)因子,在干細(xì)胞移植治療中樞神經(jīng)系統(tǒng)疾病方面是BMSCs良好的替代來源。
[Abstract]:Objective: to compare the proliferation of bone marrow mesenchymal stem cells and adipose mesenchymal stem cells. The ability of neural stem cells to induce and differentiate into neurons and other terminal cells. Methods: bone marrow mesenchymal stem cells (BMSCs) from the same SD rats were obtained. Bone marrow mesenchymal stem cells. BMSCs) and adipose mesenchymal stem cells (Adipose-derived strom cells) were isolated and cultured in vitro. Then the two kinds of mesenchymal stem cells were induced to be neurospheres and differentiated into neurons. CCK-8 was used to detect the proliferation rate of the neurons and to calculate the spheroidization rate of the two kinds of mesenchymal stem cells. The difference of induction and differentiation of mesenchymal stem cells into terminal nerve cells was compared by immunofluorescence technique. ELISA was used to detect the expression of BDNF and NT-3 neurotrophic factors. Results: the morphology of the two mesenchymal stem cells was slightly different and the two kinds of cells could be purified by subculture. ADSCs can reach the cell purity needed for induction faster than BMSCs. The growth curve shows that ADSCs can increase value faster than BMSCs. BMSCs and ADSCs are easy to be induced by stepwise method. Through the ball and differentiation into terminal nerve cells. The neuron-forming rate of ADSCs and BMSCs were 21.734 鹵3.46% and 15.62 鹵2.37, respectively, which were significantly higher than those of BMSCs (P 0.05). The positive rate of ADSCs 尾 -tubulin 鈪,
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