| Culture system in bovine mammary epithelial cell was established to study the molecularmechanism of proliferation and differentiation in bovine mammary epithelial cell. Themammary tissues from healthy bovine in the duration of lactation have been cultured in vitro.The basic medium is DMEM/F12 and 10% fetal calf serum added penicillin (0.05g/L) andstreptomycin (0.05g/L). ITS (10mL/L), 17β-E2(5mg/L), hydrocortisone (5mg/L), EGF(20ug/L) was added in different comprision. The cell biological propertied were observed andinvestigated by observing cell population doubling time and cell growth curve. Bovinemammary epithelial cell lines were obtained by culturing tissue fragment. The appearance ofthese cells was studied by light microscope. 1.The results showed that the bovine mammary epithelial cell grows well andproliferates vigorously in A2,A3 culture system. 2.The doubling time of BMEC is 47 hours. The growth curve indicate that, afterinoculation, the BMEC resort 2 days, stay in stagnate phase for 4 days, them came intologarithmic growth phase. 3.The suitable frozen medium for BMEC is DMEM/F12 added 10% DMSO and 30%FCS. The temperature should be dropped from 4℃ to -25℃ at the rate of -1℃/min,30minutes later, put the cells into liquefied nitrogen for long term preservation. 4.The BMEC were cultured by cell differentiation medium for 72 hours, and then thecasein was tested in the medium. The results indicate that we have established the bovinemammary epithelial cell line; the cell line has the function of secreting casein. 5.The fibroblast cell and the bovine mammary epithelial cells grow in different district.The bovine mammary epithelial cells were cultured in vitro through single-cell afloat, thecells formed island monolayer aggregates, than the cells propagated and formed dome-likeconstruction. 6.The cell nucleus are circular or oval which has 2-4 cores, in the culture medium withestrogen, the nucleus type changed. 7.The mammary epithelial cells can developed into many kinds cell type.
|
PDF Full Text Request