Toggle Main Menu Toggle Search

Open Access padlockePrints

Establishment of spontaneously immortalised keratinocyte lines from wild-type and mutant mice

Lookup NU author(s): Dr Julia Reichelt


Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


A considerable number of transgenic or knockout mice in which epidermal keratinocytes have been targeted die shortly after birth due to barrier defects. In this case, recovery and cultivation of keratinocytes from these animals provide an opportunity for in vitro studies. Working with isolated keratinocytes is also interesting for certain experiments which cannot be performed in live animals. Primary human keratinocytes can be kept in culture for a variable number of passages and then senesce. Immortalization can be achieved by transduction with constructs encoding viral genes. Murine keratinocytes can be kept in culture as primary cells. Naturally the numbers of cells obtained by direct isolation from mouse epidermis is restricted and sometimes not sufficient for certain biochemical analyses. To overcome this restriction some permanent murine keratinocyte lines have been generated by transfection with SV40T or HPV E6E7 genes. This is, however, not suitable if established or hypothetical biochemical links exist between these genes and the pathways or processes to be analysed in the respective experiment. We describe an easy and reproducible method of establishing permanent keratinocyte lines from spontaneously immortalised primary murine keratinocytes. This method employs co-cultivation of keratinocytes with 3T3-J2 fibroblast feeder cells for several passages during which immortalization occurs. The resulting keratinocyte lines do not only grow infinitely but, in many cases, individual lines from the same genetic background also exhibit similar growth characteristics, hence they are especially valuable for comparative studies.

Publication metadata

Author(s): Reichelt J, Haase I

Editor(s): K Turksen

Publication type: Book Chapter

Publication status: Published

Book Title: Epidermal Cells: Methods and Protocols

Year: 2009

Pages: 59-69

Online publication date: 05/09/2009

Edition: 2

Series Title: Methods in Molecular Biology

Publisher: The Humana Press Inc.

Place Published: Totowa, NJ, USA


DOI: 10.1007/978-1-60761-380-0_5

Library holdings: Search Newcastle University Library for this item

ISBN: 9781607613794