Derivation and long-term culture of cells from newt adult limbs and limb blastemas.
Notwithstanding the key importance of in vivo models for understanding patterning and cellular interactions in the regenerating tailed amphibian (salamander) limb, dissection of molecular mechanisms, as in other species, can be greatly aided by robust in vitro models. This chapter focuses on derivation and maintenance of cell lines from adult post-metamorphic salamanders and in particular cells derived from normal and regenerating limbs. We also describe a protocol for nucleofecting newt cells that can be used both to investigate the gene function in short-term studies and to establish stable cell lines.
UCL Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1EH, UK, p.ferretti@ucl.ac.uk.
Patrizia
P
Ferretti
Anoop
A
Kumar
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D017690:Cell Transplantation; D015925:Cryopreservation; D005121:Extremities / Q000502:physiology*; D012038:Regeneration; D012455:Salamandridae / Q000235:genetics / Q000502:physiology*; D013997:Time Factors; D046509:Tissue Culture Techniques; D014162:Transfection