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.

Molecular and Cellular Effects of Hydrogen Peroxide on Human Lung Cancer Cells: Potential Therapeutic Implications

“…Cells are constantly exposed to reactive oxygen species including those metabolically generated as products of aerobic respiration [ 32 , 33 ] and those originated from environmental pollutants [ 34 ]. It has been observed that hydrogen peroxide concentrations above 100 μ M are cytotoxic and genotoxic in A549 cells [ 35 ] causing DNA damage [ 36 ] and inducing the catalytic activities of DNA topoisomerase complexes [ 37 – 39 ]. Furthermore, the H 2 O 2 -induced damage could be also revealed by the oxidation of DNA bases, for instance, guanine adducts like 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxo-dG) [ 40 ].…”

Toxicogenetic study of omeprazole and the modulatory effects of retinol palmitate and ascorbic acid on Allium cepa

“…Studies of DNA damage at the chromosome level are needed in toxicogenetic studies (Sharma et. al., 2015).…”

Mesenchymal Stem Cells: An Innovative Approach in Pharmacokinetics

“…Recent study by Latifpour et al [52] report that human umbilical cord (hUCMSCs) can trans-differentiate into primordial like GCs (PGCs) when in vitro culture was performed under specific condition (bone morphogenetic protein 4 followed by retinoic acid). Hence, MSCs can be efficiently utilized to study mechanisms and pathways of different toxicants provoking etiology of many diseases, toxicity testing, evaluation of cytotoxicity and genotoxicity of pharmaceuticals and toxicity screening of chemicals that induce neurotoxicity, cardiotoxicity, hepatotoxicity, reprotoxicity, and drug discovery [53]. For example, development of anti-obesity drugs requires adipocytes that can be used to test whether any chemicals and toxicants can affect this process.…”

“…So, MSCs were shown to be capable of inducing angiogenesis and restoring the number of ovarian follicles and corpora lutea in ovaries [71]. hUCMSCs can differentiate into PGC under specific in vitro condition [53]. …”