人精原细胞的分离、纯化及培养条件的研究

发布时间：2018-03-18 17:22

本文选题：精原细胞　切入点：分离　出处：《吉林大学》2009年硕士论文　论文类型：学位论文

【摘要】： 本文通过采用联合二步酶消化法获得人生精细胞悬液,经Percoll不连续密度梯度离心,通过间隔不同时间、不同次数差异黏附法纯化人精原细胞。然后在培养液中加入不同浓度的干细胞因子、白血病抑制因子和碱性成纤维细胞生长因子后,观察细胞体外存活和增殖情况。本实验对人精原细胞的分离方法、体外培养方法进行研究,为进一步研究体内外精原干细胞诱导分化和移植打下基础,以人精原干细胞的体内外诱导分化为单倍体的细胞或精子为最终研究目标,从而为解决非梗阻性无精子症患者自己生育后代的医学难题创造条件,也为阐明人和动物生精机理提供新的思路、方法和途径。
[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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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