Comparative study of isolation, expansion and characterization of epithelial cells

Abstract

Background aimsThe 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. MethodsIn 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. ResultsThe 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. ConclusionsOur 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.