摘要
目的 观察体外培养人嗜铬细胞 (HCC)和微囊化人嗜铬细胞 (ME HCC)的儿茶酚胺(CA)及甲 脑啡肽 (M ENK)的释放功能 ,了解微囊技术对ME HCC形态和活性的影响。方法 取 6例ABO同型健康成人脑死亡的双侧肾上腺进行HCC原代培养 ,对HCC进行HE染色并行免疫细胞化学检查 ,计数酪氨酸羟化酶阳性细胞 ;用 2 %海藻酸钠经微囊技术包裹HCC并进行培养 ,在光镜下观察 ;每 4 8h更换和收集HCC和ME HCC培养液 ,- 2 0℃保存 ;采用放射免疫测定技术检测培养液及烟碱刺激试验后培养液中CA和M ENK水平。结果 (1)ME HCC形态圆整 ,体外培养生长良好 ,与HCC相似 ;(2 )HCC和ME HCC培养液中CA及M ENK水平无显著性差异 (P >0 0 5 ) ;(3)低水平烟碱刺激能提高培养液中HCC和ME -HCC释放CA及M ENK量 ,且二者CA及M ENK增加量无显著性差异 (P>0 0 5 )。结论 本包膜材料和制囊技术对HCC无损伤 ,ME HCC体外培养的活性和释放功能良好 。
Objective To compare the catecholamine (CA ) and M enkephalin(M ENK) release from human chromaffin cells (HCC) and microencapsulated HCC (ME HCC ) and investigate the effects of microencapsulation on the growth and activity of chromaffin cells Methods Adrenal glands were taken from brain death healthy adults The chromaffin cells were isolated and primarily cultured in vitro During culture the chromaffin cells were HE stained and assessed by labelling the cells with tyrosine hydroxylase monoclonal antibody The percentage of tyrosine positive cells were counted under microscope (1) The chromaffin cells were microencapsulated with 2% alginate and cultured in vitro (experimental group) HCC which were not microencapsulated were used as control The culture media of both groups were replaced every 48h and collected and stored under -20℃ for determination of CA and M ENK concentration (2) On the 6th day of culture, nicotine was added to HCC and MC HCC suspension After 30 min incubation, the suspension was centrifuged and the supernate collected and stored under -20 ℃ for determinatoin of CA and M ENK concentration with radioimmunoassay Results (1) ME HCC grew fairly well in vitro in the culture medium and was morphologically similar to HCC (2) There was no significant difference in CA and M ENK concentration in HCC and ME HCC culture media (3) CA and M ENK concentrations in supernate were increased by nicotine stimulation and there was no difference in the CA and M ENK concentration in the supernate between the two groups Conclusions Alginate and microencapsulation technique are not harmful to HCC, ME HCC has fairly good activity and release function and can be effectively used for transplantation
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2001年第7期410-413,共4页
Chinese Journal of Anesthesiology
基金
国家自然科学基金 (3 990 0 13 9)
卫生部研究基金 (98 1 12 9)资助
