Establishment And Characteristics Of Cattle Fibroblast Cell Lines

The finite fibroblast cell lines of the ear marginal tissues of Luxi cattle and Piemontese bovine were successfully constructed by direct adherent culture methods. The cell lines were analyzed according to the request for setting up cell line.The cultured cells' morpha is typical fibroblast. Through microscope the morpha diversity of fibroblast in different breeds can be observed. The cells were passaged no more than 4 generations. The cell freezing density is about 3X10Vml. Sample size of Luxi cattle is 8 and ampoules are 40; Sample size of Piemontese bovine is 2 and ampoules are 31.The cell survival rate descended after cryopreservation. This showed the cell was injured during cryopreservation and revival.The growth curve of culture cell in vitro appeared "S" shape and illustrated that cells growing included latent phase, logarithmic growth phase and stagnate phase.The result of microbial detection showed that bacteria, fungi, virus and mycoplasma did not contaminate cultured cells.The hereditary-character-stable is important for cell lines, especially for species conversation. So chromosome analysis is necessary. The result of chromosome analysis: Observing 100 cells of Luci cattle and Piemontese bovine, mostly the chromosome diploid number of Luxi cattle and Piemontese bovine is 2n=60. Morpha of chromosome is normal. The frequency of cell chromosome number to be 2n = 60 was 87%.The result showed cells still kept diploid condition and the hereditary character is stable. There is no crossed contamination between cell lines.Through analyzing two kinds of isoenzyme, MDH and LDH, different breeds has specific brands. There is no crossed contamination between cell lines.The cell lines that constructed in this study accord with the request of ATCC. We successfully constructed two kinds of cattle fibroblast cell lines and conserve genetic resource of domestic animal in cell level. And valuable materials were provided to biological studies.