An Outgrowth Culture System of Normal Human Epithelium

An In Vitro Model to Study the Coordination of Cellular Migration and Proliferation
  • Tohru Masui
Protocol
Part of the Methods In Molecular Medicine™ book series (MIMB, volume 188)

Abstract

In order to study the growth regulation of normal human epithelium, we have developed an outgrowth culture system (1). Cinematographic observation has revealed that the cells migrate and proliferate in a coordinated manner. For example, an outgrowth will expand concentrically as a cell sheet, and the cell density will be consistent between different sized outgrowths (see Fig. 1). The increment in the radius of the outgrowth stays constant (about 1.5 mm/d) and is independent of the size of the explant or donor individuals. We can easily obtain 108 cells per specimen in 3 wk. Because of its constant growth pattern, the growth history of the cultures can be easily traced, and comparable cultures can be used for further analyses (see Note 1).
Fig. 1

Macroscopic appearance of outgrowth cultured in DMMC medium supplemented with 4% FBS. Concentric circles are the records of outgrowth expansion made every 2 d.

Keywords

Migration Phenol Albumin Penicillin Trypsin 

References

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Copyright information

© Humana Press Inc. 2002

Authors and Affiliations

  • Tohru Masui
    • 1
  1. 1.Division of Genetics and MutagenesisNational Institute of Health SciencesTokyoJapan

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