Cryopreservation of Iranian Markhoz Goat Fibroblast Cells as an Endangered National Genetic Resource
Abstract The sustainable use of local animals is being eroded annually. Thus, a strategic vision for the conservation of biodiversity is of far-reaching emphasis to deal with unprecedented challenges in the local population growth process facing in the future. This study aimed to establish, characterize and cryopreserve endangered Markhoz goat (Capra hircus) fibroblast cell lines in vitro. These primary fibroblast cells were isolated from 58 Iranian Markhoz goats and individually cultured by explant technique in DMEM media supplemented with 10 % FBS. The cultured cell lines were morphologically consistent with fibroblast cells. The population doubling time for DMEM-cultured cells were 23±0.5 h. Chromosomal analysis indicated a total chromosome number of 2n = 60 with >95% frequency. Experimental assays for bacteria, fungi, yeast, and mycoplasma were reported negative. The efficiencies of VSV-G (pMDG) and lentiviral pCSGW vectors encoding fluorescent proteins showed an approximate value of 65%. Species identification for each sample was performed and confirmed correct goat cell banking without any miss- and cross-contamination. This study demonstrated the successful establishment of genetically stable fibroblast bank as a valuable genetic resource for endangered Iranian Markhoz goat breed.