Abstract

Successful somatic cell nuclear transfer aka cloning requires good quality undamaged nuclear DNA from desired cell types. In vitro culture of cells is one way of ensuring nuclear integrity. Cellular contents including nucleus gradually decompose postmortem, if not preserved within a reasonable time, leading to cell and ultimately nuclear DNA damage. The goal of this study was to determine time limits within which live and culturable cells can be obtained, after death of an animal, using sheep as a model. How long the somatic cells are alive and have potential to replicate after the animal death is not precisely known. Here we show, for the first time, that the sheep ear skin stored at 25-26°C after animal death can be cultured up to 10 days postmortem. The culture confluence is inversely correlated with increasing postmortem time interval. The cultured fibroblast-like cells have 95±5.2 % post cryopreservation cell-viability; have normal karyotype, and a comparable growth profile to that of fresh tissue derived cells. This study shows that sheep skin has potential for in vitro culture of its cells up to 10 days postmortem. Cultured cells can be successfully used for preservation of biodiversity for possible future cloning of animals.

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