【摘要】 以液氮冷冻泌乳奶牛乳腺组织为研究材料,对奶牛乳腺组织的旋转培养法与贴壁培养法进行了比较研究。乳腺组织的培养条件为:DMEM/F12培养液中含2μg/mL氢化可的松、5μg/mL转铁蛋白、10μg/mL胰岛素和10%的胎牛血清。通过台盼蓝染色观察(旋转培养法)、培养组织观察(贴壁培养法)、测定MTT二甲基噻唑二苯基四唑嗅盐活性和基因组DNA降解成为1000bp以下核苷酸片段量来比较两种方法的优缺点。结果发现,旋转培养法在120h后组织和细胞成活率都在80%以上,MTT与基因组DNA降解值显示在72h内组织活性最高;贴壁培养法组织、细胞在24h开始贴壁,到48h后完全贴壁成活,72h后组织具有明显分泌活性,MTT值与基因组DNA降解值显示,72h以后奶牛乳腺组织已表现出分裂增殖能力,组织死亡率明显下降,96h后培养组织的增殖能力与死亡率都达到稳定状态。奶牛乳腺组织贴壁和旋转两种培养方法在72h内都可以保持奶牛乳腺组织基本活性。更多还原

【Abstract】 The mammary tissue from dairy cows was cultured by the roller tube culture and stationary culture (24 well plastic culture plates). The explants were immediately incubated at 37 C (95% air; 5% CO2) in medium DMEM/F12 (4 mL volume/tube) containing 10% fetal calf serum, 2 μg/mL hydrocortisone, 10 μg/mL insulin, and 5 μg/mL transferrin. The tissue maintained the secretion activity under 72 h in roller tube culture, which was indicated by the MTT(3-(4,5-dimethylthiazol-2-y1)-2, 5-diphenyl-tetrazolium bromide) value and oligonucleosomal fraction of cellular DNA. While in stationary culture, the mammary tissue did not attach entirely to the bottom of culture plate until 24 h. The droplet of fat was secreted from the cultured mammary tissue at 72 h. In terms of the MTT value and oligonucleosomal fraction of cellular DNA, it was demonstrated that the mammary gland tissue proliferated from 72 h, and the apoptosis rate of cultured tissues was stable from 96 to 144 h. It is inferred that both the roller tube and stationary methods could maintain the basic active state of the cultured tissue before 72 h.更多还原