Establishment and Characterization of a Fibroblast Line from Sinihe Horse

For preserving the genetic resources of Sinihe Horse and exploring a new method to preserve endangered species, a Sinihe Horse fibroblast cell line was established successfully by the attachment culture. The cell line was preserved using the cryopreservation biotechniques. 48 Sinihe horse ear marginal tissue samples was used to form the cell line and has stocks of 156 cryovials, with each containing 4.5×106 cells respectively. Based on the research, Biological characteristics showed the following: During the cells proliferation, the cells were exhibited densitydependent inhibition; analysis of microbial contamination from bacteria, mycoplasma and fungi were negative; the cells population doubling time was 35.7 h; and chromosome number of the cells were 64, and the diploid frequency was higher than 80%. Isoenzyme assays of lactic and malic dehydrogenases showed no cross-contamination of this cell line with other species. Theree fluorescent proteins, pEGFP-N1, pEYFP-N1 and pDsRed1-N1, were transfected into the cells to study exogenous gene expression. The interrelated fluorescence distributed throughout the cytoplasm and nucleus 12h and 48h after transfection.The Sinihe Horse fibroblast cell line preserves the genetic resources of the Sinihe Horse at the cellular level and valuable material had been provided for genome, postgenome and somatic cloning research in this species. [Cui P, Hu PF, Li CL, Guan WJ, Ma YH. Establishment and Characterization of a Fibroblast Line from Sinihe Horse. Life Sci J 2013;10(4):1410-1416] (ISSN:1097-8135). http://www.lifesciencesite.com. 186