小鼠精原细胞分离培养及其影响因素的实验研究

发布时间：2018-05-09 06:13

本文选题：精原细胞 + 增殖　；参考：《山东大学》2005年博士论文

【摘要】：据报道,近年来男性不育的发生率有所升高,无精、少精和弱精的患者有所增加,因而对精子发生机制及其影响因素的研究也就成了生殖医学领域的一大研究热点。精子发生从精原细胞开始,经精母细胞和精子细胞,形成成熟的精子,其间经历多次有丝分裂和两次减数分裂,受神经、内分泌、旁分泌及多种细胞因子的调节,是一个极其复杂的生物学过程;对这一复杂过程及其影响因素的研究是预防和治疗男性不育和研制男性节育方法的基础性研究。目前这方面的研究多为在体研究,体外研究较少。在体研究结果显示,导致男性精子数目及质量下降的因素主要有遗传缺陷、慢性疾患、过度劳累和精神负担、环境污染、不良的饮食和生活习惯等。由于在体研究受神经、体液等许多复杂因素的影响,因而在体外培养条件下研究生精过程及其影响因素便成了必然趋势。最原始的一种精原细胞即A型精原细胞是生精干细胞,通过非对称性有丝分裂,这种细胞部分分化为B型精原细胞,另一部分则仍然保持为生精干细胞。在进行体外研究生精过程的机制及其影响因素时,最先遇到的一个问题就是如何提取和纯化培养生精干细胞。本项研究以小鼠为实验动物,首先研究了生精干细胞的提取、纯化培养的最佳方法,然后检测了温度、EGF、重组卵泡刺激素、高氧等对生精干细胞存活、代谢、增殖、凋亡等的影响。一.精原细胞的分离和纯化培养精原细胞中的A型精原细胞是一种生精干细胞,可通过非对称性细胞分裂分化为B型精原细胞并进行自我复制。目前,尚无法提取和纯化这种精原干细胞,只能提取和纯化培养含有生精干细胞的全部精原细胞。本实验筛选出了分离、培养和体外扩增精原细胞的一种比较好的方法,为生精过程及其影响因素的体外研究提供了重要的技术支持。
[Abstract]:It is reported that the incidence of male infertility has increased in recent years, and the number of patients with azoospermia, oligozoospermia and asthenospermia has increased. Therefore, the research on the mechanism of spermatogenesis and its influencing factors has become a hot topic in the field of reproductive medicine. Spermatogenesis begins with spermatogonia and forms mature spermatozoa through spermatocytes and spermatocytes, which undergo multiple mitosis and two meiosis and are regulated by nerve, endocrine, paracrine and multiple cytokines. The study of this complex process and its influencing factors is the basic research on the prevention and treatment of male infertility and the development of male birth control methods. At present, most of the researches in this field are in vivo, but few in vitro. The results showed that the main factors leading to the decline of sperm number and quality were genetic defects, chronic diseases, overwork and mental burden, environmental pollution, poor diet and living habits, etc. Since in vivo research is influenced by many complicated factors such as nerve and body fluid, it is an inevitable trend for graduate students to study the process of spermatozoa and its influencing factors under the condition of culture in vitro. The most primitive spermatogonia, type A spermatogonial cells, are spermatogenic stem cells, which differentiate into B type spermatogonial cells by asymmetric mitosis, while the other part still maintain spermatogenic stem cells. In the process of in vitro spermatogenesis and its influencing factors, one of the first problems is how to extract and purify spermatogenic stem cells. In this study, mice were used as experimental animals. The best methods for extracting and purifying spermatogenic stem cells were studied, and then the survival, metabolism and proliferation of spermatogenic stem cells were detected, such as EGF, recombinant follicle stimulating hormone, hyperoxia, etc. The effect of apoptosis, etc. I. Isolation and purification of spermatogonia Type A spermatogonia in spermatogonia is a kind of spermatogenic stem cells, which can differentiate into B spermatogonia by asymmetric cell division and self-replicate. At present, it is not possible to extract and purify spermatogonial stem cells, but only to extract and purify all spermatogonia containing spermatogenic stem cells. A better method for isolation, culture and amplification of spermatogonia in vitro was screened out in this experiment. The in vitro study on the process of spermatogenesis and its influencing factors provided important technical support.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R321.1

【引证文献】