Cryopreservation of Iranian Markhoz goat fibroblast cells as an endangered national genetic resource

چکیده مقاله

Background The continuous accessibility of local animals for sustainable use 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 extension facing in the future. This study aimed to establish and cryopreserve endangered Markhoz goat (Capra hircus) fbroblast cell lines in vitro. Methods and results These primary fbroblast cells were isolated from 58 Iranian Markhoz goats and individually cultured by explant technique in DMEM medium supplemented with 10% FBS and 2 mM L-Glutamine, in the presence of Penicillin (200 U/ml)—Streptomycin (200 mg/ml) during the frst passage number. The extracted cell lines were confrmed morphologically as fbroblast cells. The population doubling time for DMEM-cultured cells was 23±0.5 h. Chromosomal analysis indicated a total chromosome number of 2n=60 with>95% frequency. The cultured cells were checked for bacteria, fungi, yeast, and mycoplasma contaminations and the results were reported negative. The efciencies of the fuorescent protein encoded by VSV-G (pMDG) and lentiviral pCSGW vectors reported in a range of 65% value. According to the species identifcation analysis, the goat cell lines were banked and confrmed without any miss- and cross-contamination. Conclusions The signifcant issue in this paper can be concluded about the frst report of the establishment of endangered Markhoz goat cell banking inside the country. This study demonstrated the successful establishment of a genetically stable fbroblast bank as a valuable genetic resource for the endangered Iranian Markhoz goat breed