Establishment and biological characteristics of Luxi cattle fibroblast bank.

A fibroblast line from ear marginal tissue of Luxi cattle (LXCEM2/2) was successfully established by direct culturing of explants. Biological analysis showed that the population doubling time (PDT) for reviving cells was approximately 24h. Measurement of lactic dehydrogenase (LDH) and malic dehydrogenase (MDH) isoenzymes showed no cross-contamination among the cells. Karyotyping showed that the frequency of cells with chromosome number 2n=60 was 90.7-92.2%. Tests for bacteria, fungi, viruses and mycoplasma were negative. The efficiencies of expression of pEGFP-N3, pEYFP-N1 and pDsRed1-N1 were between 6.3% and 31.6% at 24h, 48h and 72h after transfer; at 24h, fluorescence was well distributed in the cytoplasm and nucleus except for some cryptomeric vesicles. Every index of the Luxi cattle cell line meets the quality control standards of the American Type Culture Collection (ATCC). Not only has the germline of this important cattle breed been preserved at the cell level, but also valuable material had been provided for genome, postgenome and somacloning research. Moreover, the establishment of this technical platform may provide both technical and theoretical support for storing the genetic resources of other animals and poultry at the cell level.