人精原細(xì)胞的分離、純化及培養(yǎng)條件的研究
本文選題:精原細(xì)胞 切入點:分離 出處:《吉林大學(xué)》2009年碩士論文 論文類型:學(xué)位論文
【摘要】: 本文通過采用聯(lián)合二步酶消化法獲得人生精細(xì)胞懸液,經(jīng)Percoll不連續(xù)密度梯度離心,通過間隔不同時間、不同次數(shù)差異黏附法純化人精原細(xì)胞。然后在培養(yǎng)液中加入不同濃度的干細(xì)胞因子、白血病抑制因子和堿性成纖維細(xì)胞生長因子后,觀察細(xì)胞體外存活和增殖情況。 本實驗對人精原細(xì)胞的分離方法、體外培養(yǎng)方法進(jìn)行研究,為進(jìn)一步研究體內(nèi)外精原干細(xì)胞誘導(dǎo)分化和移植打下基礎(chǔ),以人精原干細(xì)胞的體內(nèi)外誘導(dǎo)分化為單倍體的細(xì)胞或精子為最終研究目標(biāo),從而為解決非梗阻性無精子癥患者自己生育后代的醫(yī)學(xué)難題創(chuàng)造條件,也為闡明人和動物生精機(jī)理提供新的思路、方法和途徑。
[Abstract]:In this paper, spermatocyte suspension was obtained by combining two-step enzyme digestion method. The suspension was centrifuged by Percoll discontinuous density gradient, and the time interval was different. Human spermatogonial cells were purified by different times of different adhesion methods. After different concentrations of stem cell factor, leukemia inhibitory factor and basic fibroblast growth factor were added to the culture medium, the survival and proliferation of human spermatogonia were observed in vitro. The methods of isolation and culture of human spermatogonial cells in vitro and in vivo were studied in this experiment, which laid a foundation for further study on differentiation and transplantation of spermatogonial stem cells in vivo and in vitro. Human spermatogonial stem cells were induced to differentiate into haploid cells or spermatozoa in vitro and in vivo, so as to create conditions for solving the medical problems of non-obstructive azoospermia patients to reproduce their own offspring. It also provides new ideas, methods and approaches for elucidating the mechanism of spermatogenesis in humans and animals.
【學(xué)位授予單位】:吉林大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2009
【分類號】:R329
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