Brown D G, Willington M A, Findlay I, Muggleton-Harris A L
MRC Experimental Embryology and Teratology Unit, St. George's Hospital Medical School, Tooting, London, United Kingdom.
In Vitro Cell Dev Biol. 1992 Nov-Dec;28A(11-12):773-8. doi: 10.1007/BF02631066.
Cultured mouse embryonic stem (ES) cells are used for both in vitro and in vivo studies. The uncommitted pluripotent cells provide a model system with which to study cellular differentiation and development; they can also be used as vectors to carry specific mutations into the mouse genome by homologous recombination. To ensure successful integration into the germ line, competent totipotent diploid ES cell lines are selected using a cell injection bioassay that is both time consuming and technically demanding. The prolonged in vitro culture of rapidly dividing ES cells can lead to accumulated changes and chromosomal abnormalities that will compromise the biological function and abrogate germ line transmission of chimeric mice carrying novel genetic mutations. Such in vitro conditions will vary between individual laboratories; for example, differences in the serums used for maintenance. Using a number of different criteria we attempt in this paper to define the parameters that we found to be key factors for optimization of the biological potential of established ES cell lines. The successful integration into the germ line is dependant on acquiring or deriving a competent totipotent mouse ES diploid cell line. In this paper parameters and criteria are defined which we found to be key factors for the optimization of the biological potential of established ES cell lines.
培养的小鼠胚胎干细胞（ES细胞）用于体外和体内研究。未分化的多能细胞提供了一个研究细胞分化和发育的模型系统；它们还可以作为载体，通过同源重组将特定突变导入小鼠基因组。为确保成功整合到种系中，使用既耗时又技术要求高的细胞注射生物测定法来选择有能力的全能二倍体ES细胞系。快速分裂的ES细胞长时间体外培养会导致累积变化和染色体异常，这将损害生物学功能并消除携带新基因突变的嵌合小鼠的种系传递。这种体外条件在不同实验室之间会有所不同；例如，用于维持培养的血清存在差异。在本文中，我们使用多种不同标准尝试定义我们发现的对优化已建立的ES细胞系生物学潜能至关重要的参数。成功整合到种系中取决于获得或衍生出有能力的全能小鼠ES二倍体细胞系。本文定义了一些参数和标准，我们发现这些是优化已建立的ES细胞系生物学潜能的关键因素。
