A simple non-invasive protocol to establish primary cell lines from tail and toe explants for cytogenetic studies in Australian dragon lizards (Squamata: Agamidae)

被引：24

作者：

Ezaz, Tariq ^{[1
,2
]}

O'Meally, Denis ^{[2
]}

Quinn, Alexander E. ^{[1
]}

Sarre, Stephen D. ^{[1
]}

Georges, Arthur ^{[1
]}

Graves, Jennifer A. Marshall ^{[2
]}

机构：

[1] Univ Canberra, Inst Appl Ecol, Canberra, ACT 2601, Australia

[2] Australian Natl Univ, Comparat Genom Grp, Res Sch Biol Sci, Canberra, ACT 2601, Australia

来源：

CYTOTECHNOLOGY | 2008年 / 58卷 / 03期

基金：

澳大利亚研究理事会;

关键词：

Reptiles; Lizards; Cell lines; Tail; Toe; Non-invasive; Chromosomes;

D O I：

10.1007/s10616-009-9182-3

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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 A degrees 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.

引用

页码：135 / 139

页数：5