Cell and Tissue Banking

, Volume 14, Issue 3, pp 423–435 | Cite as

Characterization of cultured epithelial cells using a novel technique not requiring enzymatic digestion for subculturing

  • Antonio Peramo
  • Stephen E. Feinberg
  • Cynthia L. Marcelo
Original Paper


Our laboratory had developed a methodology to expand epithelial cells in culture by growing keratinocyte monolayers, under large volumes of medium that produces large numbers of keratinocytes that leave the monolayer and move into suspension. The cells have been defined as epithelial Pop Up Keratinocytes or ePUKs cells and appear to be highly suitable for clinical applications. In this publication we extend the characterization of the cells with a detailed analysis of the capabilities of the monolayer of a single culture flask to produce, over time, ePUK cells. The cells were characterized using standard epithelial markers for proliferation and differentiation. Analysis of morphology of the monolayer formed and total number of cells produced is presented for a variety of human epithelial cell strains. These keratinocytes provide an additional controlled human cell system for investigation of the mechanisms regulating epithelia cell growth and differentiation and since they are produced in large numbers, they are highly suitable for use in epithelial cell banking.


Keratinocyte ePUK Epithelia cell Cell culture Oral mucosa 

Supplementary material

10561_2012_9343_MOESM1_ESM.doc (67 kb)
Supplementary material 1 (DOC 67 kb)
10561_2012_9343_MOESM2_ESM.tif (181 kb)
Figure S1 Comparison of cell production for the pairs ePUK0-ePUK1 for different cell strains. ePUK production of frozen strains show a substantial decrease in the ePUK1 flasks. In fresh cells, neonatal strains maintain a similar production from the beginning, but oral ePUK1 cells grow more slowly (TIFF 180 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Antonio Peramo
    • 1
    • 2
  • Stephen E. Feinberg
    • 2
  • Cynthia L. Marcelo
    • 1
  1. 1.Department of SurgeryUniversity of MichiganAnn ArborUSA
  2. 2.Department of Oral and Maxillofacial SurgeryUniversity of MichiganAnn ArborUSA

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