Establishment and biological characteristics of Piedmontese cattle fibroblast line

Abstract

A fibroblast line was successfully established from ear marginal tissues of Piedmontese cattle by direct culturing of explants. Biological analysis showed that the population doubling time (PDT) for reviving cells was approximately 24 h. The average viability of the cells was 96.6% before freezing and 92.7% after thawing. Isoenzyme polymorphorism of lactic dehydrogenase (LDH) and malic dehydrogenase (MDH) ruled out cross-contamination among cell lines. Karyotyping showed that the proportion of cells with chromosome number 2n = 60 was above 92%. Tests for bacteria, fungi, viruses and mycoplasmas were negative. The transfection efficiencies of pEGFP-N3, pEYFP-N1 and pDsRed1-N1 were between 7.1 and 30.8%; fluorescences were homogenously distributed throughout cytoplasm and nucleus except in some cryptomeric vesicles. Every index of the Piedmontese cattle cell line met the quality control standards of the American Type Culture Collection (ATCC).

Key words: Piedmontese cattle, fibroblast line, biological characterization.