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Development of an organotypic stem cell model for the study of human embryonic palatal fusion

Cynthia J Wolf1, David G Belair1, Carrie M Becker2

  • 1Toxicity Assessment Division, National Health and Environmental Effects Research Laboratories, Office of Research and Development, US EPA Research Triangle Park, North Carolina.

Birth Defects Research
|October 23, 2018
PubMed

Insights

A new 3D organoid model accurately mimics human palatal fusion. Human epidermal growth factor (hEGF) was found to impede fusion and increase proliferation in this model, offering insights into cleft palate development.

Area of Science:

  • Biomedical Engineering
  • Developmental Biology
  • Stem Cell Research

Background:

  • Cleft palate (CP) is a common congenital birth defect affecting 1 in 1,000 births globally.
  • Palatal fusion failure, specifically the epithelial seam between shelves, is a key mechanism causing CP.
  • Epidermal growth factor (EGF) influences palate development, potentially hindering fusion.

Purpose of the Study:

  • To develop and validate a 3D organotypic model of human palatal fusion.
  • To investigate the effects of human EGF (hEGF) on the proliferation and fusion of embryonic palatal shelves in vitro.

Main Methods:

  • A 3D organotypic model was created using human mesenchymal stem cells (hMSCs) and progenitor epithelial keratinocytes (hPEKs).
  • hMSCs were differentiated into an osteogenic lineage and formed spheroids, which were then coated with hPEKs to create heterotypic spheroids (organoids).
  • Organoids were cultured with or without hEGF to assess cell proliferation and fusion dynamics.

Main Results:

  • Osteogenic differentiation of hMSCs peaked by Day 13.
  • Exposure to hEGF delayed organoid fusion at 12 and 18 hours.
  • hEGF at 4 ng/ml increased proliferation in the organoids, with proliferation also observed in hPEKs alone.

Conclusions:

  • The developed 3D organoid model effectively replicates human palatal fusion morphology and response to hEGF.
  • This model serves as a valuable tool for studying the mechanisms underlying cleft palate formation and potential therapeutic interventions.
Abstract

Keywords:
cleft palatefusionhuman stem cellsorganotypic modelsspheroid

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