两种不同方法培养大鼠肾上腺细胞的比较  

Comparison of two different methods for rat adrenocortical cells culture

作　　者：方立[1] 张杰[1] 杜贤进[1] 李又空[1] 彭建平[1] 

机构地区：[1]武汉大学人民医院泌尿外科,430060

出　　处：《中华实验外科杂志》2009年第10期1272-1274,I0003,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30471734）

摘　　要：目的观察胶原凝胶三维培养对肾上腺细胞形态和功能的影响。方法分离SD大鼠乳鼠肾上腺细胞，于自制鼠尾Ⅰ型胶原凝胶中培养14d，与传统平面培养的肾上腺细胞相对比，比较两者在细胞形态、细胞增殖率、蛋白表达和分泌功能方面的差异。结果两种培养方式下，肾上腺细胞均能表达其特异性细胞色素氧化酶CYP11B1和CYP11B2。原代培养8d后，三维胶原凝胶培养大鼠肾上腺细胞的增殖率显著高于平面培养，至第14天，胶原凝胶中培养的肾上腺细胞增殖率为16．6％，而传统平面培养的仅为2．9％。胶原凝胶中培养的肾上腺细胞分泌功能维持时间更长，第14天其培养液中皮质酮和醛固酮浓度分别为（278．33±38．20）μg/L和（133．85±38．20）ng／L，而相应的平面培养肾上腺细胞分泌皮质酮和醛固酮浓度分别为（160．35±28．12）μg／L和（74．03±12．12）ng／L，差异有统计学意义（P〈0．05）。结论胶原凝胶中三维培养肾上腺细胞更有利于细胞功能的维持。Objective To investigate a novel 3-dimensional cell culture model in collagen gel, and its influence on morphology and function of adrenoeortieal cells. Methods Adrenoeortieal cells of neonate rats were isolated, and then cultured in self-made collagen type I gel for 14 days. This 3-dimensional cell culture model was compared with the classic plane culture model on the morphology, cell growth rate, protein expression and secretory functions. Results Adrenocortical cells expressed specific cytochrome P450 ：CYP11 B1 and CYP11 B2 under both cuhure models. At the 8th day after primary culture ,the growth rate of adrenocortical cells in 3-dimensional cell culture model was higher than the classic plane culture model significantly. At the 14th day, the growth rate in 3-dimensional cell culture model and the classic plane culture model was 16.6% and 2.9% respectively. The secretory functions of adrenocortical cells were more sustainable in 3-dimensional collagen gel. At the 14th day, corticosterone and aldosterone levels in the medium of 3-dimensional cell culture model were （278.33 ± 38.20） μg/L and （133.85 ± 38.20） ng/L, while those were （ 160.35 ± 28.12 ） μg,/L and （ 74.03 ± 12.12 ） ng/L in the medium of plane culture model, with the differences being statistically significant （ P 〈 0.05 ）. Conclusion Threedimensional culture in collagen gel is favorable for the maintenance of adrenocortical cells functions.

关 键 词：Ⅰ型胶原 肾上腺细胞 培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]