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Related Experiment Video

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Methods for Differentiating hiPSCs into Vascular Smooth Muscle Cells.

Mei-Lan Li1, Jiesi Luo2,3, Matthew W Ellis2,3,4

  • 1Biological and Biomedical Sciences Program, Yale University, New Haven, CT, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 30, 2021
PubMed
Summary

Human induced pluripotent stem cells (hiPSCs) offer a promising source for vascular smooth muscle cells (VSMCs) in tissue-engineered vascular grafts (TEVGs). This chapter details methods for generating mature VSMCs from hiPSCs for advanced cardiovascular therapies.

Keywords:
Embryoid body differentiation methodHuman induced pluripotent stem cellsMonolayer differentiation methodTissue-engineered vascular graftsVascular bypassingVascular smooth muscle cells

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Area of Science:

  • Regenerative Medicine
  • Stem Cell Biology
  • Vascular Tissue Engineering

Background:

  • Generating functional vascular tissues for tissue-engineered vascular grafts (TEVGs) faces challenges in sourcing suitable vascular cells.
  • Human pluripotent stem cells, particularly human induced pluripotent stem cells (hiPSCs), provide a scalable and patient-specific source for vascular smooth muscle cells (VSMCs).
  • Autologous hiPSC-derived VSMCs offer immunocompatibility, crucial for preventing graft rejection in TEVG applications.

Purpose of the Study:

  • To detail three distinct methods for differentiating hiPSCs into mature, functional VSMCs suitable for TEVG engineering.
  • To highlight the potential of hiPSC-derived VSMCs in advancing cell-based therapies for cardiovascular diseases.

Main Methods:

  • Embryoid body (EB)-based differentiation for large-scale, robust production of mature hiPSC-derived VSMCs, mimicking early embryonic development.
  • Chemically defined differentiation systems to generate embryological origin-specific hiPSC-derived VSMCs from defined germ layers (neuroectoderm, mesoderm) for disease modeling.
  • Monolayer culture on extracellular matrix proteins combined with pulsatile flow to promote the secretion of organized elastic fibers by hiPSC-derived VSMCs.

Main Results:

  • Successful differentiation of hiPSCs into structurally and functionally mature VSMCs using diverse methodologies.
  • Demonstration of large-scale production capabilities for hiPSC-derived VSMCs via EB formation.
  • Establishment of methods for generating lineage-specific VSMCs and enhancing extracellular matrix organization.

Conclusions:

  • hiPSCs represent a vital cell source for generating patient-specific VSMCs, overcoming limitations in current TEVG technology.
  • The presented differentiation strategies provide a foundation for developing advanced TEVGs and cellular therapies for vascular repair.
  • The unlimited supply of hiPSC-derived VSMCs heralds a new era in treating cardiovascular diseases through regenerative approaches.