Comparative study of isolation, expansion and characterization of epithelial cells

Cytotherapy. 2017 Feb;19(2):263-271. doi: 10.1016/j.jcyt.2016.10.008. Epub 2016 Nov 25.

Abstract

Background aims: The human epithelial cells (EPCs) have been identified as the essential element for the regeneration of skin construct for burns, wounds and various tissue engineer-based products.

Methods: In this study, the isolation, expansion and characterization of EPCs from various sources such as juvenile foreskin (JSK), buccal mucosa (BM), penile skin (PS) and urothelium (UR) in serum-free and xeno-free EpiLife media were evaluated.

Results: The growth kinetics study revealed that EPCs from JSK and BM had notably higher growth rates compared with the others. Overall, the EPCs from all sources retained basic morphological characteristics and the functional characteristics such as Pan Cytokeratin (AE1/AE3). In addition, the cryopreservation stability of EPCs was accessed for post-thaw viability and found to be greater than 80% at 1 year of storage, but demonstrated reduced cell recovery (51%) at the second year in fetal bovine serum-free cryopreservation media.

Conclusions: Our result suggests that the EPCs from four cell sources can be grown in feeder-free, serum-free and xeno-free systems using commercially available EpiLife medium without losing epithelial cell characteristics even after passage 4. However, its suitability for clinical application must be accessed by preclinical and clinical studies.

Keywords: cell therapy; epithelial cell source; epithelial cells; growth kinetics; tissue engineering.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Aged
  • Cell Culture Techniques / methods*
  • Cell Proliferation*
  • Cell Separation / methods*
  • Cell Survival
  • Cells, Cultured
  • Child
  • Child, Preschool
  • Epithelial Cells / cytology*
  • Epithelial Cells / physiology*
  • Freezing / adverse effects
  • Humans
  • Immunophenotyping
  • Infant
  • Male
  • Middle Aged