CYTOTOXICITY TEST USING BLASTOCYST-DERIVED EUPLOID EMBRYONAL STEM-CELLS - A NEW APPROACH TO INVITRO TERATOGENESIS SCREENING
CYTOTOXICITY TEST USING BLASTOCYST-DERIVED EUPLOID EMBRYONAL STEM-CELLS - A NEW APPROACH TO INVITRO TERATOGENESIS SCREENING
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DOI：
10.1016/0890-6238(91)90111-r
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发表时间：
1991-01-01
影响因子：
3.3
通讯作者：
SPIELMANN, H
中科院分区：
文献类型：
--
作者：
LASCHINSKI, G;VOGEL, R;SPIELMANN, H
为了建立一种用于致畸性试验的哺乳动物体外系统，在来自小鼠囊胚的多能整倍体胚胎干细胞（ESC）中评价了外源性物质的细胞毒性。二甲基噻唑-二苯基溴化四氮唑（MTT）试验是ESC细胞毒性测定的最合适的试验系统。仅选择不需要代谢活化的化合物从数据库中进行测试，以验证Smith等人的体外致畸试验。将ESC获得的结果与第14天小鼠胚胎成纤维细胞的相应数据进行比较，以检测未分化和分化细胞之间的灵敏度差异。ESC对已知致畸剂的敏感性高于成纤维细胞培养物，这允许在哺乳动物系统中计算“成体”细胞（分化的成纤维细胞）与胚胎细胞（未分化的ESC）的敏感性比率，类似于水螅试验。虽然在我们的系统中，一些外源性物质被归类为假阴性，但ESC细胞毒性试验有望成为畸形学中一种新的体外筛选试验。
To develop a mammalian in vitro system for teratogenicity testing, cytotoxicity of xenobiotics was evaluated in pluripotent euploid embryonal stem cells (ESC) derived from mouse blastocysts. The dimethylthiazol-diphenyl tetrazolium bromide (MTT) assay was the most appropriate test system for cytotoxicity determinations with ESC. Only compounds that do not require metabolic activation were selected for testing from the database for validation of in vitro teratogenesis assays by Smith et al. Results obtained with ESC were compared to corresponding data from fibroblasts from day-14 mouse embryos to detect differences in sensitivity between undifferentiated and differentiated cells. ESC showed a higher sensitivity to known teratogens than fibroblast cultures, which allows calculation of a sensitivity ratio of "adult" cells (differentiated fibroblasts) to embryonal cells (undifferentiated ESC) in a mammalian system similar to the hydra assay. Although some xenobiotics had to be classified as false negatives in our system, the ESC cytotoxicity assay holds promise as a new in vitro screening assay in teratology.