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Cellular characterization and successful transfection of serially subcultured normal human esophageal keratinocytes
Author(s) -
Compton Carolyn C.,
Warland Gretchen,
Nakagawa Hiroshi,
Opitz Oliver G.,
Rustgi Anil K.
Publication year - 1998
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199811)177:2<274::aid-jcp9>3.0.co;2-k
Subject(s) - rous sarcoma virus , transfection , involucrin , biology , cell culture , keratinocyte , microbiology and biotechnology , fibroblast , 3t3 cells , cellular differentiation , stratified squamous epithelium , cell growth , in vitro , epithelium , immunology , virus , gene , genetics
Background In vitro cell culture models can provide unique insights into squamous epithelial proliferation, differentiation, and neoplastic transformation. Cultures of human esophageal keratinocytes could be advantageous for the study of these processes. Methods: Normal human esophageal keratinocytes were cultivated on 3T3 fibroblast feeder layers in vitro and expanded through four serial subcultivations. Confluent tertiary cultures were analyzed by morphological and immunohistochemical techniques to define their basic properties. The ability to transiently transfect cultured esophageal epithelium was tested using a Rous sarcoma virus‐luciferase reporter gene by the calcium phosphate and lipofection methods. Results: Postconfluent cultures displayed a predominantly basal cell phenotype with limited stratification, widespread expression of keratins 5 and 14, and production of attachment specialization proteins such as α6β4 integrin and collagen VII. Terminal differentiation markers (involucrin and transglutaminase) were prematurely expressed. The cells expressed growth factors important in proliferation and differentiation, such as transforming growth factor‐β and interleukin‐1β.Tertiary cultures were successfully transiently transfected with a Rous sarcoma virus‐luciferase reporter gene construct. Conclusion: Normal human esophageal cells can be serially passaged through extended numbers of cell generations and transfected by standard methods. This in vitro system may be useful in the study of fundamental cellular processes governing proliferation and differentiation in the esophageal epithelium. J. Cell. Physiol. 177:274–281, 1998. © 1998 Wiley‐Liss, Inc.