title: Human mesenchymal stem cells as a novel platform for simultaneous evaluation of cytotoxicity and genotoxicity of pharmaceuticals.
abstract: The in vitro micronucleus test is a well-known test for the screening of genotoxic compounds. However until now, most studies have been performed on either human peripheral lymphocytes or established cancer cell lines. This study provides human mesenchymal stem cells as an alternative to the conventional micronucleus test. We grew umbilical cord mesenchymal stem cells (UC-MSCs) on coverslips eliminating the cumbersome technique involving hypotonic treatment, fixation and preparing smears required for suspension culture (lymphocytes). The background frequency of nuclear blebs and micronuclei in UC-MSCs was found to be 7±5, in lymphocytes 16±3.5 and 9±3 and that for A549 cell line was 65±5 and 15±5 per 1000 cells, respectively, suggesting differences in the repair mechanism of normal and cancer cell lines. We inspected the cytotoxic and genotoxic effects of two known mutagens, mitomycin-C and hydrogen peroxide (H2O2), on UC-MSCs, lymphocytes and A549 cells. Treatment with mitomycin-C and H2O2 demonstrated drastic differences in the degree of cytotoxicity and genotoxicity suggesting a constitutional difference between normal and cancer cells. In addition we tested two solvents, dimethyl sulfoxide (DMSO) and ethanol, and two drugs, metformin and rapamycin. DMSO above 1% was found to be cytotoxic and genotoxic, whereas ethanol at same concentration was neither cytotoxic nor genotoxic indicating the minimal non-toxic level of the solvents. This study thus offers UC-MSCs as a better substitute to peripheral lymphocytes and cancer cell lines for high throughput screening of compounds and reducing the animal studies.
authors.affiliation: School of Regenerative Medicine, Manipal University, GKVK Post, Bellary Road, Yelahanka, Bangalore, Karnataka 560065, India,Biochemistry and Stem Cell Research, National Institute of Nutrition (NIN), Jamai-Osmania PO, Hyderabad, Andhra Pradesh 500007, India andDepartment of Toxicology, International Institute of Biotechnology and Toxicology (IIBAT), Padappai, Kancheepuram, Tamilnadu 601301, India.
authors.forename: Shikha
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Biochemistry and Stem Cell Research, National Institute of Nutrition (NIN), Jamai-Osmania PO, Hyderabad, Andhra Pradesh 500007, India and.
authors.forename: Vijayalakshmi
authors.initials: V
authors.lastname: Venkatesan
authors.affiliation: Department of Toxicology, International Institute of Biotechnology and Toxicology (IIBAT), Padappai, Kancheepuram, Tamilnadu 601301, India.
authors.forename: Balakrishna Murthy
authors.initials: BM
authors.lastname: Prakhya
authors.affiliation: School of Regenerative Medicine, Manipal University, GKVK Post, Bellary Road, Yelahanka, Bangalore, Karnataka 560065, India,Biochemistry and Stem Cell Research, National Institute of Nutrition (NIN), Jamai-Osmania PO, Hyderabad, Andhra Pradesh 500007, India andDepartment of Toxicology, International Institute of Biotechnology and Toxicology (IIBAT), Padappai, Kancheepuram, Tamilnadu 601301, India rr.bhonde@manipal.edu.
authors.forename: Ramesh
authors.initials: R
authors.lastname: Bhonde
mesh_terms: D000328:Adult; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016922:Cellular Senescence; D004249:DNA Damage; D004121:Dimethyl Sulfoxide / Q000633:toxicity; D004353:Drug Evaluation, Preclinical / Q000379:methods; D000431:Ethanol / Q000633:toxicity; D005260:Female; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000502:physiology; D008687:Metformin / Q000633:toxicity; D015162:Micronucleus Tests; D009153:Mutagens / Q000633:toxicity*; D020123:Sirolimus / Q000633:toxicity
chemical_list: D009153:Mutagens; D000431:Ethanol; D008687:Metformin; D020123:Sirolimus; D004121:Dimethyl Sulfoxide