Amnion epithelial cell isolation and characterization for clinical use

Curr Protoc Stem Cell Biol. 2010 Apr;Chapter 1:Unit 1E.6. doi: 10.1002/9780470151808.sc01e06s13.

Abstract

Human amnion epithelial cells (hAECs) are a heterologous population positive for stem cell markers; they display multilineage differentiation potential, differentiating into cells of the endoderm (liver, lung epithelium), mesoderm (bone, fat), and ectoderm (neural cells). They have a low immunogenic profile and possess potent immunosuppressive properties. Hence, hAECs may be a valuable source of cells for cell therapy. This unit describes an efficient and effective method of hAEC isolation, culture, and cryopreservation that is animal product-free and in accordance with current guidelines on preparation of cells for clinical use. Cells isolated using this method were characterized after 5 passages by analysis of karyotype, cell cycle distribution, and changes in telomere length. The differentiation potential of hAECs isolated using this animal product-free method was demonstrated by differentiation into lineages of the three primary germ layers and expression of lineage-specific markers analyzed by PCR, immunocytochemistry, and histology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amnion / cytology
  • Amnion / physiology*
  • Animals
  • Cell Cycle
  • Cell Differentiation
  • Cell Separation / methods*
  • Cryopreservation / methods*
  • Epithelial Cells / cytology*
  • Flow Cytometry / methods
  • Humans
  • Immunohistochemistry / methods
  • Immunosuppressive Agents / pharmacology
  • Karyotyping / methods
  • Mice
  • Polymerase Chain Reaction
  • Telomere / ultrastructure

Substances

  • Immunosuppressive Agents