Differentiated Fibroblastic Progenies of Human Embryonic Stem Cells for Toxicology Screening

摘要

Immortalized cell lines and live animal models are commonly used for cytotoxicity screeningnof biomedical devices and materials. However, these assays poorly reflect human physiologynand have numerous other disadvantages. An alternative may be to utilize differentiated fibroblasticnprogenies of human embryonic stem cells (hESC) for in vitro toxicology screening.nThese were generated through random spontaneous differentiation within standard culturenmedia, over several passages. The cytotoxic response of the differentiated hESC fibroblasticnprogenies (pH9) to mitomycin C was observed to be not only very similar to the L929 cell line,nbut was, in fact, more sensitive. At an initial seeding density of 1000 cells/well (0.33 cm2), the proliferationnindex was observed to decrease 19.0% from 1.638 to 1.326 for the L929 cell line, asnthe dosage of mitomycin C was gradually increased from 0 to 1.54 u0002g/mL. By contrast, pH9ndisplayed a corresponding 40.5% drop in proliferation index from 3.713 to 2.209. At a highernseeding density of 2000 cells/well (0.33 cm2), the proliferation index was observed to decreasen27.0% from 1.213 to 0.885 for the L929 cell line, whereas pH9 displayed a corresponding 43.7%ndrop in proliferation index from 3.711 to 2.091. Hence, it is apparent that pH9 exhibited a morensensitive dose–response to mitomycin C compared to L929, which could be advantageous forncytotoxicity screening assays. Additionally, this study also demonstrated that a highly purifiednand well-defined phenotypic population of differentiated hESC progenies is not necessarynfor high reproducibility and accuracy in cytotoxic response.