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Attempt at conserving the genetic resources of Hu sheep by fibroblast line cryopreservation

Author:
Li, Xiangchen, Hu, Pengfei, He, Xiaohong, Lu, Taofeng, Guan, Weijun, Ma, Yuehui
Source:
Journal of applied animal research 2014 v.42 no.3 pp. 352-355
ISSN:
0974-1844
Subject:
acrocentric chromosomes, autosomes, cross contamination, cryopreservation, ears, fibroblasts, fluorescent proteins, freezing, genes, isozymes, karyotyping, microbial contamination, plasmids, sex chromosomes, sheep, sheep breeds, thawing, viability, China
Abstract:
Hu sheep is one of the most distinctive local sheep breeds in China, while investigations concerning its cell biology and molecular genetics are still scarce. To preserve the valuable genetic resources of the Hu sheep, we established a Hu sheep fibroblast line from 35 sheeps' ear marginal tissues using explant culture and cryopreservation techniques, and its biological characteristics were identified, including cell morphology, growth curve, karyotype, isoenzyme polymorphism, and expression of exogenous fluorescent genes. The results showed that the viability of the fibroblasts before freezing was 96.54 ± 3.22% and 92.36 ± 2.77% after thawing, and there was no microbial contamination and cross-contamination; the growth curve of Hu sheep ear marginal fibroblasts displayed an obvious “S” shape; all somatic chromosomes were acrocentric autosomes, and only the two sex chromosomes (XY) were submetacentric; the normal number of chromosomes (2 n = 54) was observed in 96.56 ± 2.89% of the cells; and the transfer efficiencies of four fluorescent proteins could achieve 37.4% with an optimized ratio of plasmid and lipofectamine. It could be concluded that this study has not only provided the biological characteristics of Hu sheep at cellular level, but also makes a valuable contribution to the preservation of the genetic resources of the Hu sheep.
Agid:
1225176