A fibroblast line from kindey tissue of Hereford cattle was established successfully by direct culture of explants and biology cryopreservation techniques. The cell line contained 101 tubes of frozen cells from 34 primary kidney samples. Biological analysis showed that the cells were morphologically consistent with fibroblasts and the growth curve was sigmoidal with a Population Doubling Time (PDT) of 35 h.The average viability of the cells was 95.8% before freezing and 93.4% after thawing. Cross-contamination among cell lines was excluded by isoenzyme analysis of Lactate Dehydrogenase (LDH) and Malate Dehydrogenase (MDH). The frequency of cells having the diploid chromosome number (60) was 97.5%. Detection of bacteria, fungi, viruses and mycoplasmas was verified negative. At 24, 48 and 72 h after transfection, the expression efficiency of fluorescent protein genes (pEGFP-N3, pEYFP-N1 and pDsRed1-N1) were between 18.6~32%; The fluorescence could be observed well-distributed in cytoplasm and nucleus in addition to some cryptomere vesicles at 12 h after transfection.
展开▼
