论文部分内容阅读
目前,嵌合体技术已成为胚胎干细胞(ES细胞)和诱导多能性干细胞(iPS细胞)研究不可或缺的一部分,嵌合体小鼠模型主要用于基因功能、调控机制、致病机理等各方面的研究。要安全可靠地进行ES细胞和iPS细胞的各种应用性研究,首先需要验证细胞在体内向各种组织定向分化的能力,特别是生殖系传递的能力。而高效制备多能性干细胞生殖系传递的嵌合体小鼠,需要在分子水平、细胞水平、动物个体水平以及制备方法等各方面全面综合地分析考虑,才能达到实验设计的最优化。制备嵌合体小鼠主要有注射法和聚集法2种方法,各有利弊,在整个制备过程中,需要考虑不同发育阶段以及不同小鼠品系胚胎的分化潜能、选用远交系和杂交系的胚胎作为受体,同时要考虑ES细胞的遗传背景,选择低代次的远交系和杂交系的ES细胞或者iPS细胞,以及优选KSR培养体系。对于操作者,需根据实际情况,全方位地综合考虑,从而制定出合理的操作方案。
At present, the chimera technology has become an indispensable part of embryonic stem cell (ES cell) and induced pluripotent stem cell (iPS cell) research. The chimeric mouse model is mainly used for gene function, regulatory mechanism, pathogenic mechanism and other aspects Research. To safely and reliably perform various applied studies on ES cells and iPS cells, it is first necessary to verify the ability of the cells to differentiate into various tissues in vivo, in particular, the germline transmission. However, efficient preparation of chimeric mice delivered by pluripotent stem cell germ plasm requires comprehensive and comprehensive consideration and analysis at molecular level, cell level, animal’s individual level, and preparation method, so as to achieve the optimization of experimental design. Preparation of chimeric mice are mainly injection and aggregation of the two methods, each has its advantages and disadvantages, throughout the preparation process, need to consider different stages of development and differentiation potential of different mouse embryos, selection of inbred lines and hybrid embryos As an acceptor, the genetic background of ES cells should be taken into consideration at the same time. ES cells or iPS cells of low-generation inbred and hybrid lines, and preferably KSR culture system, should be selected. For the operator, according to the actual situation, comprehensive consideration in all directions, in order to work out a reasonable operating plan.