胚胎小体制备新方法的建立及小鼠胚胎干细胞体外分化形成新生血管的实验研究

发布时间：2018-01-12 02:23

本文关键词：胚胎小体制备新方法的建立及小鼠胚胎干细胞体外分化形成新生血管的实验研究　出处：《大连医科大学》2006年硕士论文　论文类型：学位论文

【摘要】： 研究背景和目的胚胎干细胞(embryonic stem cells, ES cells)是从胚胎早期内细胞团(inner cell mass, ICM)分离得到的全能干细胞,在体外培养时如去除饲养层细胞和白血病抑制因子( leukemia inhibitory factor, LIF)等维持干细胞未分化状态的培养条件,ES细胞可自发形成具有内、中、外三胚层结构的胚胎小体(embryoid bodies, EBs)。作为ES细胞体外分化的理想模型,EBs可以再现胚胎早期发育过程中的谱系特异性分化程序及基因时空表达模式的诸多方面,因此被广泛用于研究胚层间相互诱导、器官空腔化形成等胚胎早期发育事件。此外,在由ES细胞向特定组织细胞诱导分化的过程中,往往需要经过EBs阶段的培养,以完成ES细胞分化的启动。由此可见,EBs制备已成为干细胞研究领域不可或缺的工具。目前常用的EBs制备方法,如悬滴培养法、悬浮培养法、甲基纤维素半固体培养法等,均存在着操作繁琐、EBs产生数量少、大小和分化阶段不一致等缺点,直接影响了相关研究的可行性及结果的可靠性。为改进EBs制备过程中存在的上述问题,本实验通过采用先贴壁培养ES细胞3 d,再悬浮培养的策略,探讨建立一种制备EBs的新方法。近年来随着癌症和心脑血管疾病的增多,人们越来越关注血管形成机制的研究。充分认识这一过程,将对恶性肿瘤及缺
[Abstract]:Background and purpose of the study Embryonic stem cells (es cells) are derived from inner cell mass in early embryo. In vitro culture, such as the removal of feeder layer cells and leukemia inhibitory factor. Embryo embryoid bodies with inner, mesoderm and ectodermal structure can be formed spontaneously under the condition of maintaining the undifferentiated state of stem cells, such as LIFs. As an ideal model for es cell differentiation in vitro, EBs can reproduce the pedigree specific differentiation process and gene expression pattern in early embryonic development. Therefore, it has been widely used to study early embryonic development events such as interdermal induction, organ cavity formation and so on. In addition, in the process of inducing differentiation from es cells to specific tissue cells. It is often necessary to culture es cells through EBs stage to complete the initiation of es cell differentiation. Thus, the preparation of EBs has become an indispensable tool in the field of stem cell research. At present, EBs preparation methods are commonly used. For example, suspension culture method, suspension culture method, methyl cellulose semi-solid culture method and so on, there are some shortcomings such as less production of EBs in operation, inconsistent size and differentiation stage, and so on. In order to improve the above problems in the process of EBs preparation, we adopted the strategy of culture es cells for 3 days and then suspension culture. To establish a new method for preparing EBs. In recent years, with the increase of cancer and cardiovascular and cerebrovascular diseases, people pay more and more attention to the study of angiogenesis mechanism.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329

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