The Validated Embryonic Stem Cell Test with Murine Embryonic Stem Cells.
The Validated Embryonic Stem Cell Test with Murine Embryonic Stem Cells.
复制标题
使用小鼠胚胎干细胞进行经过验证的胚胎干细胞测试。
DOI：
10.1007/978-1-4939-7883-0_4
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
ZurNieden,NicoleI
中科院分区：
文献类型：
--
作者：
Walker,LaurenM;McClelland-Descalzo,DarcieLV;ZurNieden,NicoleI
出生缺陷是美国婴儿死亡的主要原因，但大多数情况的原因尚不清楚。虽然在大多数情况下怀疑是遗传和环境因素共同作用的结果，但关于产前接触许多常见化学物质对胎儿造成的健康风险的信息却很少。因此，开发和完善能够准确预测化合物胚胎毒性的程序对于减少出生缺陷婴儿的数量非常重要。胚胎干细胞试验 (EST) 是一种利用胚胎和成体细胞的细胞毒性比较以及分化抑制来预测所测试化合物的胚胎毒性的程序。由于使用现有细胞系，EST 极大地减少了毒性测试中对动物测试对象的需求。此外，由于 EST 使用分化抑制作为终点，因此在可测试的并发症范围内用途极其广泛。本章解释了使用经过验证的胚胎干细胞测试和传统心肌细胞分化终点的程序。该协议包括常规干细胞培养、心肌细胞分化程序以及利用分子终点评估化合物胚胎毒性的方法的讨论。
Birth defects are the leading cause of infant mortality in the USA, yet the causes of most of these conditions are unknown. While a combination of genetic and environmental factors are suspected in most cases, little information exists about the health risks that prenatal exposure to many common chemicals poses for the fetus. Thus, development and refinement of procedures that can accurately predict embryotoxicity of compounds is important for curtailing the number of infants born with birth defects. The embryonic stem cell test (EST) is a procedure that utilizes comparison of cytotoxicity in embryonic and adult cells and inhibition of differentiation to predict embryotoxicity of compounds tested. Because of its use of existing cell lines, the EST dramatically reduces the need for animal test subjects in toxicity testing. In addition, because of its use of inhibition of differentiation as an endpoint, the EST is extremely versatile in the range of complications it can test for. In this chapter, procedures for use of the validated embryonic stem cell test with the traditional cardiomyocyte differentiation endpoint are explained. The protocol includes discussion of routine stem cell culture, the cardiomyocyte differentiation procedure, and methods for utilization of molecular endpoints for assessing embryotoxicity of compounds.