Chapter 5. Culture of Human Cervical Epithelial Cells

  1. R. Ian Freshney and
  2. Mary G. Freshney
  1. Margaret A. Stanley

Published Online: 28 APR 2002

DOI: 10.1002/0471221201.ch5

Culture of Epithelial Cells, Second Edition

Culture of Epithelial Cells, Second Edition

How to Cite

Stanley, M. A. (2002) Culture of Human Cervical Epithelial Cells, in Culture of Epithelial Cells, Second Edition (eds R. I. Freshney and M. G. Freshney), John Wiley & Sons, Inc., New York, USA. doi: 10.1002/0471221201.ch5

Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

Author Information

  1. Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK

Publication History

  1. Published Online: 28 APR 2002
  2. Published Print: 12 APR 2002

Book Series:

  1. Culture of Specialized Cells

Book Series Editors:

  1. R. Ian Freshney

Series Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

ISBN Information

Print ISBN: 9780471401216

Online ISBN: 9780471221203

SEARCH

Keywords:

  • cervical epithelium;
  • epithelial cell culture;
  • 3T3 feeder layers;
  • serum-free medium;
  • KGM;
  • MCDB 153;
  • cytokeratin;
  • CIN;
  • cervical carcinoma;
  • involucrin;
  • collagen rafts;
  • BPV immortalization

Summary

The chapter starts with an anatomical and histological description of the uterine cervix, and an historical review of attempts at culture. Reagent preparation and protocols are provided for the culture of endo- and ectocervical keratinocytes on 3T3 feeder layers following trypsin disaggregation of mechanically separated epithelium. The preparation of the feeder layer is described and instructions given for subculture with feeder layers. A separate protocol is given for culture with serum-free KGM (MCDB 153). Culture cervical intraepithelial neoplasms (CIN) and carcinoma is described using a primary explantation without trypsinization. Characterization of cervical epithelia cell cultures has been achieved using cytokeratin and involucrin expression. Conditions for maximal expression of differentiation are provided using collagen rafts. Protocols for immortalization with BPV are also described.