大鼠肾上腺细胞体外培养观察及鉴定

发布时间：2018-06-07 10:12

本文选题：肾上腺细胞 + 细胞培养　；参考：《赣南医学院学报》2010年06期

【摘要】：目的:探讨大鼠肾上腺细胞体外培养时的生长特征及分泌功能,从而为寻找到肾上腺细胞最佳细胞移植时限提供实验依据。方法:通过肾上腺细胞的体外分组原代培养,取不同培养时间(0、2、4、6、7、10 d)的肾上腺细胞,倒置相差显微镜下定性观察肾上腺细胞的形态,细胞贴壁生长及其分布情况;定量检测肾上腺细胞增殖情况及其存活率;酶联免疫分析法测定培养液中皮质酮的浓度,放射免疫分析法测定培养液中皮质醇和醛固酮的浓度。比较分析肾上腺细胞分泌功能的动态变化。结果:正常大鼠肾上腺细胞体外原代培养48 h后,在显微镜下见肾上腺细胞贴壁,细胞增大,胞体呈圆形或多角形,胞体大,胞浆透亮,4 d后生长旺盛,数量增多,细胞成活率达95.94%,2周后细胞出现皱缩和破碎,开始衰老。体外培养的肾上腺细胞培养液中,皮质醇浓度随时间的延长而增加,但第4天以后增加速度缓慢,进入一个平台期。皮质酮、醛固酮浓度在第2、4、6、7 d逐渐上升,第7 d达峰值.随后第10 d浓度开始下降,与高峰值相比,有显著差异(P0.01),而皮质醇略升高。结论:体外培养的大鼠肾上腺细胞具有分泌皮质激素的功能;大鼠体内所含糖皮质激素主要为皮质酮,皮质醇虽然有变化,但其含量非常低。
[Abstract]:Aim: to investigate the growth characteristics and secretory function of rat adrenal cells in vitro, so as to provide experimental basis for finding the best time of adrenal cell transplantation. Methods: the adrenal cells were cultured in vitro, and the adrenal cells were collected for 710 days. The morphologies, adherent growth and distribution of adrenal cells were observed qualitatively under inverted phase contrast microscope. The proliferation and survival rate of adrenal cells were measured quantitatively, the concentrations of cortisol and aldosterone in culture medium were determined by enzyme linked immunosorbent assay (Elisa), and the concentrations of cortisol and aldosterone in culture medium were determined by radioimmunoassay. To compare and analyze the dynamic changes of adrenal cell secretory function. Results: after primary culture in vitro for 48 h, the adrenal cells adhered to the wall under microscope, the cells were enlarged, the cell body was round or polygonal, the cell body was large, and the cytoplasm of the adrenal cells grew exuberantly and the number increased after 4 days. The cell survival rate reached 95.94g and 2 weeks later, the cells crumpled and broken, and began to senescence. In the adrenal cells cultured in vitro, the concentration of cortisol increased with the increase of time, but after 4 days, the increase rate was slow and entered a plateau phase. The concentration of corticosterone and aldosterone gradually increased on the 7th day and reached the peak on the 7th day. On the 10th day, the concentration began to decrease, there was a significant difference between the peak value and the peak value, but the cortisol increased slightly. Conclusion: in vitro cultured rat adrenal cells have the function of secreting corticosteroids and the main glucocorticoids in rats are corticosterone and the content of cortisol is very low.
【作者单位】：南昌大学研究生院医学部;赣南医学院第一附属医院泌尿外科;赣南医学院基础医学院;
【基金】：江西省卫生系统学术和技术带头人培养对象科技计划项目
【分类号】：R329

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