Cellular characterization and successful transfection of serially subcultured normal human esophageal keratinocytes

Carolyn C. Compton, Gretchen Warland, Hiroshi Nakagawa, Oliver G. Opitz, Anil K. Rustgi

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)274-281
Number of pages8
JournalJournal of Cellular Physiology
Volume177
Issue number2
DOIs
StatePublished - Nov 1998
Externally publishedYes

Fingerprint

Luciferases
Keratinocytes
Viruses
Cell culture
Transfection
Rous sarcoma virus
Genes
Keratin-14
Keratin-5
Feeder Cells
Reporter Genes
Transglutaminases
Differentiation Antigens
Cell growth
Transforming Growth Factors
Fibroblasts
Epithelium
Interleukin-1
Integrins
Intercellular Signaling Peptides and Proteins

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Cellular characterization and successful transfection of serially subcultured normal human esophageal keratinocytes. / Compton, Carolyn C.; Warland, Gretchen; Nakagawa, Hiroshi; Opitz, Oliver G.; Rustgi, Anil K.

In: Journal of Cellular Physiology, Vol. 177, No. 2, 11.1998, p. 274-281.

Research output: Contribution to journalArticle

Compton, Carolyn C. ; Warland, Gretchen ; Nakagawa, Hiroshi ; Opitz, Oliver G. ; Rustgi, Anil K. / Cellular characterization and successful transfection of serially subcultured normal human esophageal keratinocytes. In: Journal of Cellular Physiology. 1998 ; Vol. 177, No. 2. pp. 274-281.
@article{9d736929c4304f5fb4075297ac67e225,
title = "Cellular characterization and successful transfection of serially subcultured normal human esophageal keratinocytes",
abstract = "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.",
author = "Compton, {Carolyn C.} and Gretchen Warland and Hiroshi Nakagawa and Opitz, {Oliver G.} and Rustgi, {Anil K.}",
year = "1998",
month = "11",
doi = "10.1002/(SICI)1097-4652(199811)177:2<274::AID-JCP9>3.0.CO;2-K",
language = "English (US)",
volume = "177",
pages = "274--281",
journal = "Journal of Cellular Physiology",
issn = "0021-9541",
publisher = "Wiley-Liss Inc.",
number = "2",

}

TY - JOUR

T1 - Cellular characterization and successful transfection of serially subcultured normal human esophageal keratinocytes

AU - Compton, Carolyn C.

AU - Warland, Gretchen

AU - Nakagawa, Hiroshi

AU - Opitz, Oliver G.

AU - Rustgi, Anil K.

PY - 1998/11

Y1 - 1998/11

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0031707461&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031707461&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1097-4652(199811)177:2<274::AID-JCP9>3.0.CO;2-K

DO - 10.1002/(SICI)1097-4652(199811)177:2<274::AID-JCP9>3.0.CO;2-K

M3 - Article

C2 - 9766524

AN - SCOPUS:0031707461

VL - 177

SP - 274

EP - 281

JO - Journal of Cellular Physiology

JF - Journal of Cellular Physiology

SN - 0021-9541

IS - 2

ER -