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

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Abstract

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.

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Acknowledgments

This work was supported by grant R01AM-26009 to CLM.

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Author notes

  1. Antonio Peramo

    Present address: Department of Oral and Maxillofacial Surgery, University of Michigan, Ann Arbor, MI, 48109, USA

Authors and Affiliations

  1. Department of Surgery, University of Michigan, Ann Arbor, MI, 48109, USA

    Antonio Peramo & Cynthia L. Marcelo

  2. Department of Oral and Maxillofacial Surgery, University of Michigan, Ann Arbor, MI, 48109, USA

    Stephen E. Feinberg

Authors
  1. Antonio Peramo
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  2. Stephen E. Feinberg
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  3. Cynthia L. Marcelo
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Correspondence to Antonio Peramo.

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Supplementary material 1 (DOC 67 kb)

10561_2012_9343_MOESM2_ESM.tif

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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Peramo, A., Feinberg, S.E. & Marcelo, C.L. Characterization of cultured epithelial cells using a novel technique not requiring enzymatic digestion for subculturing. Cell Tissue Bank 14, 423–435 (2013). https://doi.org/10.1007/s10561-012-9343-z

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  • DOI: https://doi.org/10.1007/s10561-012-9343-z

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