Evaluation of developmental toxicity using undifferentiated human embryonic stem cells

Abstract

An embryonic stem cell test (EST) has been developed to evaluate the embryotoxic potential of chemicals with an in vitro system. In the present study, novel methods to screen toxic chemicals during the developmental process were evaluated using undifferentiated human embryonic stem (hES) cells. By using surface marker antigens (SSEA-4, TRA-1-60 and TRA-1-81), we confirmed undifferentiated conditions of the used hES cells by immunocytochemistry. We assessed the developmental toxicity of embryotoxic chemicals, 5-fluorouracil, indomethacin and non-embryotoxic penicillin G in different concentrations for up to 7 days. While expressions of the surface markers were not significantly affected, the embryotoxic chemicals influenced their response to pluripotent ES cell markers, such as OCT-4, NANOG, endothelin receptor type B (EDNRB), secreted frizzled related protein 2 (SFRP2), teratocarcinoma-derived growth factor 1 (TDGF1), and phosphatase and tensin homolog (PTEN). Most of the pluripotent ES cell markers were down-regulated in a dose-dependent manner after treatment with embryotoxic chemicals. After treatment with 5-fluorouracil, indomethacin and penicillin G, we observed a remarkable convergence in the degree of up-regulation of development, cell cycle and apoptosis-related genes by gene expression profiles using an Affymetrix GeneChips. Taken together, these results suggest that embryotoxic chemicals have cytotoxic effects, and modulate the expression of ES cell markers as well as development-, cell cycle- and apoptosis-related genes that have pivotal roles in undifferentiated hES cells. Therefore, we suggest that hES cells may be useful for testing the toxic effects of chemicals that could impact the embryonic developmental stage.

Coupling switch of P2Y-IP3 receptors mediates differential Ca2+ signaling in human embryonic stem cells and derived cardiovascular progenitor cells

“…This in vitro differentiation system provides unique tools for the study of early cardiovascular development [5], drug screening [6,7], cytotoxicity testing [8,9], and cardiac regenerative therapy [10,11]. However, the physiological and pharmacological properties of hESCs and CVPCs, such as the Ca 2+ signaling and the response to various modulators, are largely unknown.…”

An engineered mouse embryonic stem cell model with survivin as a molecular marker and EGFP as the reporter for high throughput screening of embryotoxic chemicals in vitro

“…The gene expression profiles of mouse and human ES cells have been proven to be distinct in, for example, several differentiation markers (Liu, Yin, & Faiola, 2017;Tandon & Jyoti, 2012;Wobus & Loser, 2011), which may lead to completely different responses to the test chemicals in molecular approaches (Yamane et al, 2016). Several recent studies have thus investigated the potential application of human embryonic stem cells (hESCs), associated with a variety of molecular endpoints, to predict the developmental toxicity in vitro (Ehashi, Suzuki, Ando, Sumida, & Saito, 2014;Hong & Jeung, 2013;Jung et al, 2014;Sandstrom et al, 2017). However, in addition to ethical issues, using hESCs for drug screening not only requires complex culturing conditions but also is costly and difficult to implement in the HT manner (Fang, Zhi, Yu, Lynch, & Jia, 2018;Rezvanfar, Hodjat, & Abdollahi, 2016).…”

Relationship between TLR4 and MCP2 expression levels and habitual abortion

“…The abnormal expression of MCP2 will directly lead to embryonic diapauses or might affect the embryo implantation and growth, indirectly causing abortion. Therefore, the study of the changes in expression of MCP2 has important significance in illustrating the pathogenesis of habitual abortion (Park et al, 2014;Pérez-Garijo and Steller, 2014;Hsuuw and Chan, 2015;Jung et al, 2015). TLR4 is one of the important human innate immune receptors.…”