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Induced Pluripotent Stem Cells01:06

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Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
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EPS and iPS Cells in Disease Research01:21

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Embryonic and induced pluripotent stem cells are excellent models for disease research because of their ability to self-renew and differentiate into most cell types. Somatic cells from a patient are isolated and reprogrammed into induced pluripotent stem cells or iPSCs. These iPSCs are later differentiated into the desired cell type, which mirrors the diseased cell of the patient. In this way, disease models have been created for investigating diseases such as Down syndrome, type I diabetes,...
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Embryonic Stem Cells00:57

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Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
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Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation
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The Human Induced Pluripotent Stem Cell Test as an Alternative Method for Embryotoxicity Testing.

Saskia Galanjuk1, Etta Zühr1, Arif Dönmez1

  • 1IUF-Leibniz Research Institute for Environmental Medicine, 40225 Duesseldorf, Germany.

International Journal of Molecular Sciences
|March 25, 2022
PubMed
Summary

This study introduces a new human induced pluripotent stem cell (hiPSC) test to assess chemical embryotoxicity, offering an ethical alternative to animal testing for improved human safety.

Keywords:
CardioVisioncardiomyocytesdevelopmental toxicityembryotoxicityhiPS TesthiPSCin vitrovideo analyses

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Area of Science:

  • Developmental toxicology
  • Stem cell biology
  • In vitro toxicology

Background:

  • Regulatory toxicology heavily relies on animal studies for assessing reproductive and developmental toxicity.
  • There is a critical need for alternative in vitro methods to align with the 3R principles (Replacement, Reduction, Refinement).
  • Human cell-based models are valuable for overcoming species differences and enhancing human safety assessments.

Purpose of the Study:

  • To develop and characterize a fetal bovine serum (FBS)-free in vitro test using human induced pluripotent stem cells (hiPSCs) to evaluate the embryotoxic potential of substances.
  • To establish a reliable human-based assay for developmental toxicity screening.

Main Methods:

  • Human induced pluripotent stem cells (hiPSCs) were cultured for 10 days to differentiate into beating cardiomyocytes.
  • A fetal bovine serum (FBS)-free culture condition was established for the hiPSC test (hiPS Test).
  • Terminal endpoint evaluations included cell viability, qPCR analysis, and video analysis of cardiomyocyte beating frequency and area.

Main Results:

  • The hiPS Test successfully differentiated hiPSCs into beating cardiomyocytes.
  • The assay correctly identified embryotoxic and non-embryotoxic controls: 5-Fluorouracil (5-FU) as positive and Penicillin G (PenG) as negative.
  • Key endpoints like cell viability, gene expression, and cardiomyocyte beating were measured.

Conclusions:

  • The developed hiPS Test is a promising in vitro method for evaluating substance-induced embryotoxicity using human cells.
  • Further compound screening is necessary to define the assay's applicability domain and confirm its suitability for detecting adverse effects on embryonic development.
  • This human cell-based approach offers a potential alternative to animal testing in regulatory toxicology.