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Related Experiment Video

Updated: Jun 6, 2025

Generating iPS Cells from MEFS through Forced Expression of Sox-2, Oct-4, c-Myc, and Klf4
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Mouse Embryonic Fibroblasts Reprogramming to Induced Pluripotent Stem Cells by T3.

Ana Montero-Pedrazuela1, Constanza Contreras-Jurado2,3,4

  • 1Instituto de Investigaciones Biomédicas Sols-Morreale, Consejo Superior de Investigaciones Científicas (CSIC), Universidad Autónoma de Madrid (UAM), Madrid, Spain. amontero@iib.uam.es.

Methods in Molecular Biology (Clifton, N.J.)
|November 23, 2024
PubMed
Summary

This study details a method to improve induced pluripotent stem cell (iPSC) generation using triiodo-L-thyronine (T3) hormone. This enhanced reprogramming protocol boosts iPSC production and aids in studying cell plasticity for regenerative medicine.

Keywords:
Mouse embryonic fibroblastsPluripotencyQuantitative real-time PCRReprogrammingThyroid hormoneYamanaka factorsYamanaka factorsiPSC colony formation

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Area of Science:

  • Stem cell biology
  • Molecular biology
  • Endocrinology

Background:

  • Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) using specific transcription factors (Yamanaka factors).
  • Optimizing reprogramming efficiency is crucial for advancing regenerative medicine and disease modeling.

Purpose of the Study:

  • To present a protocol for enhancing induced pluripotent stem cell (iPSC) generation from mouse embryonic fibroblasts (MEFs) using triiodo-L-thyronine (T3).
  • To describe methods for analyzing iPSC pluripotency, including alkaline phosphatase staining and gene expression profiling.

Main Methods:

  • Reprogramming of MEFs using retroviral delivery of Yamanaka factors, with the addition of triiodo-L-thyronine (T3).
  • Colony staining for alkaline phosphatase activity to identify pluripotent stem cells.
  • Quantitative real-time PCR (qPCR) to analyze endogenous pluripotency gene expression in expanded iPSC colonies.

Main Results:

  • Triiodo-L-thyronine (T3) supplementation enhances the efficiency of induced pluripotent stem cell (iPSC) generation.
  • Established iPSC colonies exhibit alkaline phosphatase activity, confirming pluripotency.
  • Gene expression analysis confirms the endogenous expression of key pluripotency markers in T3-enhanced iPSCs.

Conclusions:

  • Integrating triiodo-L-thyronine (T3) into reprogramming protocols offers a significant improvement in induced pluripotent stem cell (iPSC) production.
  • This enhanced method facilitates the generation of functional iPSCs, supporting research in cell plasticity, disease modeling, and regenerative therapies.