摘要
【目的】研究在幼仓鼠肾细胞（BHK-21）悬浮培养过程中,不同浓度氨对其生长代谢的影响。【方法】用添加0,1.8,2.5,5.5,8.0和15.0 mmol/L氨的培养基培养BHK-21细胞,每隔12 h取样1次,用细胞计数仪进行细胞计数,测定细胞活力,采用AO/EB双染色法和Annexin V-FITC/PI流式细胞仪2种方法检测细胞的凋亡情况,同时测定细胞培养过程中葡萄糖、氨和乳酸浓度的变化。【结果】在BHK-21细胞连续培养过程中,当氨的添加浓度低于1.8 mmol/L时,其对细胞的生长几乎不产生抑制作用,细胞活性、细胞总数与对照组差异不显著,未出现细胞聚集成团现象;当氨添加浓度达到15.0 mmol/L时,BHK-21细胞直接进入死亡期;当氨的添加浓度为2.5~8.0 mmol/L时,氨对细胞生长的抑制作用由弱变强,这种抑制作用在细胞培养至24 h开始出现。另外,随着氨添加浓度的增加,葡萄糖的消耗量减少,细胞代谢产生的乳酸和氨降低。【结论】氨对悬浮培养的BHK-21细胞的生长和代谢有显著影响。
【Objective】 The study was carried out to research the effects of ammonion in different concentrations during the culture on BHK-21 suspension cells.【Method】 The media of 0,1.8,2.5,5.5,8.0 and 15.0 mmol/L ammonion were used to culture the BHK-21 suspension cells.Sampling was taken every 12 h,cells were counted and vitality was determined by the cell count instrument.Double AO/EB dyeing and Annexin V-FITC/PI flow cytometric analysis were used to detect cell apoptosis,concentration of glucose,ammonia,and lactic acid in the process of cell metabolism.【Result】 The growth and metabolism of BHK-21 cells weren＇t influenced after 1.8 mmol/L of ammonia was added.Cell activity and number were not significantly different with the control group,and the phenomenon of the cell conglobation didn＇t appear;When the concentration reached above 15.0 mmol/L,the growth stopped.The ammonion at a concentration between 5.5 and 8.0 mmol/L,showed obvious inhibiting effect on the growth and metabolism of BHK-21 cells at 24 h.Along with the addition of ammonia,the consumption of glucose decreased,the same with product of lactic acid and ammonion.【Conclusion】 Ammonia showed significant influence on the growth and metabolism of BHK-21 cells.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2011年第2期28-32,共5页
Journal of Northwest A&F University(Natural Science Edition)
基金
中农威特生物科技股份有限公司资助项目(YF-20080001)
