|
Objective:To conduct the primarily culture of rat adrenocortical cells, and to observe the characteristics of growth and secretion in cell curture in different phase which will help offer experiment evidence for searching the most suitable time of adrenal cell transplantation.Methods:Adrenal glands were removed from two week old Wistar rats under sterile conditions, then the cortex was separated and shredded. The cells were primarily cultured in DMEM medium containing with 20% fetal bovine serum. The growth process and morphological features of the cells were observed dynamically under the inverted phase contrast microscope, meanwhile, the survival rate of the cells were detected by staining with trypan blue. At different culture times, the concentration of corticosterone was determined by enzyme immunoassay, corticosteroid and aldosterone by radio immunoassay.Result:After normal adrenocortical cells were primarily cultured 48 hours, cells were attached to the bottom of culture dishes and volume began to augment, most of which were round or polygonal shaped, and cell body was big and cytoplasm was lucency. Furthermore, there are many grana intracytoplasm and it’s bulk is comparatively regular, cells grew prosperitily, cell quantity increased, which became confluent and the survival rate of the cells reached to 95.94% four days after plating. With elongation of culturing time, cells became fat and fusiform shape gradually, grana intracytoplasm grew downwards by degrees. After 2 weeks, cells gradually became shrunken, broken and senesced.In adrenocortical cell culture in vitro, corticosteroid measurements showed that corticosteroid concentration was increasing with the time extending, in contrast to gradually decreased and then entered into a plateau phase after four days. The concentrations of corticosterone and aldosterone were increasing gradually in the 2th, 4th,6th or 7th day. It reached to a peak in the 7th day, and then degraded in the 10th day. Compared to the peak, there was a significant difference (P<0.01), but corticosteroid was steped up a little.Conclusion:Normal rat’s adrenocortical cells cultured primarily in vitro have the function of secreting adrenocortical hormone. Glucocorticoids in rat’s vivo is mainly corticosterone, notwithstanding, cortisol was either changed, it’s content is very low. The characteristics of growth and secretion in rats adrenocortical cell is correlated to culturing time.
|