Differentiated fibroblastic progenies of human embryonic stem cells for toxicology screening.
Immortalized cell lines and live animal models are commonly used for cytotoxicity screening of biomedical devices and materials. However, these assays poorly reflect human physiology and have numerous other disadvantages. An alternative may be to utilize differentiated fibroblastic progenies of human embryonic stem cells (hESC) for in vitro toxicology screening. These were generated through random spontaneous differentiation within standard culture media, over several passages. The cytotoxic response of the differentiated hESC fibroblastic progenies (pH9) to mitomycin C was observed to be not only very similar to the L929 cell line, but was, in fact, more sensitive. At an initial seeding density of 1000 cells/well (0.33 cm(2)), the proliferation index was observed to decrease 19.0% from 1.638 to 1.326 for the L929 cell line, as the dosage of mitomycin C was gradually increased from 0 to 1.54 microg/mL. By contrast, pH9 displayed a corresponding 40.5% drop in proliferation index from 3.713 to 2.209. At a higher seeding density of 2000 cells/well (0.33 cm(2)), the proliferation index was observed to decrease 27.0% from 1.213 to 0.885 for the L929 cell line, whereas pH9 displayed a corresponding 43.7% drop in proliferation index from 3.711 to 2.091. Hence, it is apparent that pH9 exhibited a more sensitive dose-response to mitomycin C compared to L929, which could be advantageous for cytotoxicity screening assays. Additionally, this study also demonstrated that a highly purified and well-defined phenotypic population of differentiated hESC progenies is not necessary for high reproducibility and accuracy in cytotoxic response.
Cao
Tong
T
Department of Oral-Maxillo Facial Surgery, Faculty of Dentistry, National University of Singapore, Singapore. omscaot@nus.edu.sg
Lu
Kai
K
Fu
Xin
X
Heng
Boon Chin
BC
D000465:Algorithms; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D054697:Cytostatic Agents / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D020869:Gene Expression Profiling; D006801:Humans; D016685:Mitomycin / Q000494:pharmacology; D018675:Toxicity Tests / Q000379:methods*
D015415:Biomarkers; D054697:Cytostatic Agents; D016685:Mitomycin