Chapter 4 Primary Fibroblast Cell Culture

Abstract

This chapter describes a way to culture stable primary fibroblast cell lines from adult zebrafish caudal fins in a standard tissue culture medium with bovine serum. These primary fibroblasts retain many features of nontransformed cells such as eudiploidy, contact inhibition, and surface adhesion. One of the stable fibroblast lines, AB9, derived from an AB strain, exhibits contact inhibition and surface-growth dependency. A karyotype analysis of these cells showed eudiploid chromosome count (50 chromosomes/diploid cell). The cells have been maintained in continuous culture for over 35 passages without a change in characteristics. A continuous source of stable cultured fish cells offers a variety of applications. In addition, high-quality zebrafish genomic DNA can be easily obtained from these cells. These primary fibroblasts retain the same genetic polymorphism as the original donor fish when a variety of zebrafish genes are analyzed. Therefore, cultured cells derived from mutant zebrafish could prove useful in positional cloning and the identification of mutant genes in much the same way that cultured fibroblasts are in human genetic disease research. In vitro applications of these cultured fibroblasts have been discussed in the chapter.