摘要
[目的]在高原环境条件下初步建立体外肝细胞长期培养的一种方法。[方法]将体外分离的大鼠肝细胞以3×105/孔接种于用胶原包被的12孔塑料培养板内,总体培养时间为30 d,动态观察培养细胞形态、细胞数量及培养上清中总蛋白、白蛋白和尿素水平变化。[结果]细胞接种4 h开始贴壁,8 h开始增殖,6 d生长满瓶,形成均匀的单层;4 d细胞数量明显增多,8 d最多,达35.2×105/孔,然后细胞数量渐减少;14 d培养细胞胞浆内开始出现颗粒状物质和空泡,20 d培养细胞开始出现细胞脱壁现象,培养液中漂浮细胞增多;培养上清中总蛋白、白蛋白和尿素水平均呈现显著升高,其达峰值时间分别为10、8和10 d,然后渐降低,培养结束时仍然高于培养2 d时的水平。[结论]在高原环境条件下,体外长期培养肝细胞生长、增殖及其生物学功能良好,可以应用于高原肝脏生理及病理生理等相关方面的研究。
[ Objective] To establish a new method of long-term cultivation of rat hepatocytes in plateau. [ Methods] The hepatocytes isolated from rat liver were inoculated at the culture plate coated with collagen and were cultured for 30 days. The changes of cell shape, cell amount and the levels of total protein, albumin and urea in supernatants of rat hepatocytes culture were observed. [Results] After cultured for 4 hours, the hepatocytes began to adhibit to the wall of culture plate; 8 hours later, the hepatocytes began to proliferate, some of them were binuclear; and 6 days later, the bottom of culture plate had been suffused with growing hepatocytes. The cell number began to increase 4 days later, and amounted to the maxinmm of 35.2×10^5 per hole after cultured for 8 days, then declined gradually. In the 20th days, the cultured cell began to be detached from the wall of culture plate, the amount of floating cells in culture superoatants increased. The levels of total protein, albumin and urea in rat hepatocytes culture supernatants increased remarkably and reached the peak on the days of 10, 8 and 10, respectively, besides, the level of them at the end of cultivation was still higher than that on the second day of cultivation. [Conclusions] The long term cultivation of rat hepatocytes in plateau can be applied in the study on liver physiology, pathophysiology and relevant aspects.
出处
《现代预防医学》
CAS
北大核心
2007年第20期3998-3999,共2页
Modern Preventive Medicine
基金
西藏自治区十五重点课题(2001-99)
