Primary cell lines were established from cultures of tail and toe clips of five species of Australian dragon lizards: Tympanocryptis pinguicolla, Tympanocryptis sp., Ctenophorus fordi, Amphibolurus norrisi and Pogona vitticeps. The start of exponential cell growth ranged from 1 to 5 weeks. Cultures from all specimens had fibroblastic morphology. Cell lines were propagated continuously up to ten passages, cryopreserved and recovered successfully. We found no reduction in cell viability after short term (<6 months) storage at −80 °C. Mitotic metaphase chromosomes were harvested from these cell lines and used in differential staining, banding and fluorescent in situ hybridisation. Cell lines maintained normal diploidy in all species. This study reports a simple non-invasive method for establishing primary cell lines from Australian dragon lizards without sacrifice. The method is likely to be applicable to a range of species. Such cell lines provide a virtually unlimited source of material for cytogenetic, evolutionary and genomic studies.
Reptiles Lizards Cell lines Tail Toe Non-invasive Chromosomes
Cogger HG (2000) Reptiles and amphibians of Australia, 6th edn. Reed New Holland, SydneyGoogle Scholar
Ezaz T, Quinn AE, Georges A, Sarre SD, O’Meally D, Graves JAM (2009) Molecular marker suggests rapid changes of sex-determining mechanisms in Australian dragon lizards. Chromosome Res. doi:10.1007/s10577-008-9019-5Google Scholar
Freshney IR (2006) Culture of animal cells: manual of basic technique, 4th edn. Wiley-Liss, Inc., New YorkGoogle Scholar
Harlow PS (2004) Temperature-dependent sex determination in lizards. In: Valenzuela N, Lance VA (eds) Temperature dependent sex determination in vertebrates. Smithsonian Institution, Washington DC, pp 11–20Google Scholar
Hugall AF, Foster R, Hutchinson M, Lee MSY (2008) Phylogeny of Australasian agamid lizards based on nuclear and mitochondrial genes: implications for morphological evolution and biogeography. Biol J Linn Soc 93:343–358CrossRefGoogle Scholar
Mansell JM, Elliott RJ, Gaskin JM (1989) Initiation and ultrastructure of a reptilian fibroblast cell line obtained from cutaneous fibropapillomas of the green turtle, Chelonia mydas. In Vitro Cell Dev Biol 25:1062–1064. doi:10.1007/BF02624142CrossRefGoogle Scholar