微囊化牛肾上腺嗜铬细胞多巴胺含量的检测  

THE DYNAMIC CHANGES OF DOPAMINE SECRETED BYMICROCAPSULATED BOVINE CHROMAFFIN CELLS IN VITRO

作　　者：张进禄[1] 张京钟[1] 蔡青[1] 鲁强[1] 赵春礼[1] 段春礼[1] 刘玉军[1] 薛毅珑[2] 徐群渊[1] 

机构地区：[1]首都医科大学北京神经科学研究所北京市神经损伤修复研究重点实验室,北京100054 [2]解放军总医院临床基础研究所,北京100853

出　　处：《解剖学报》2004年第5期489-492,共4页Acta Anatomica Sinica

基　　金：国家重点基金研究规划--脑功能和脑重大疾病的基础研究资助项目 (G19990 5 40 0 8) ;国家自然科学基金国际合作资助项目 ;北京市跨世纪人才工程资助项目

摘　　要：目的　检测体外培养微囊化牛嗜铬细胞分泌多巴胺的动态变化 ,为用微囊化肾上腺髓质细胞移植治疗帕金森病提供实验依据。　方法　常规培养牛肾上腺髓质嗜铬细胞 ,用人工APA微囊对之微囊化后再培养 2 0d ,高效液相色谱 (HPLC)动态检测多巴胺含量 ,并分别在光镜及电镜下观察微囊内牛嗜铬细胞的形态变化。　结果牛嗜铬细胞在微囊内生长良好 ,在培养液内可持续检测到多巴胺。第 8d左右多巴胺的分泌达到高峰 ,随后逐渐下降 ,14d后分泌量处于一种恒定的状态。　结论　微囊化牛嗜铬细胞分泌的多巴胺可以通透微囊 ,可以考虑用来进行异种细胞移植治疗帕金森病。Objective To explore the possibility of intracerebral grafting of microcapsulated bovine chromaffin cells in Parkinson disease(PD),the dynamic changes of dopamine secreted by microcapsulated bovine chromaffin cells was observed. Methods The fresh bovine adrenal medulla was dissociated with retrograde perfusion technique to obtain chromaffin cells and purified by Percoll density gradient centrifugation.The cells were then cultured in DMEM/F12 containing 10% FCS.After microcapsulation with alginate-polylysine-alginate(APA) semipermeable membrane,the concentration of dopamine secreted by these cells was determined by HPLC every two days and the morphologic changes were observed under light microscope and electron microscope respectively. Results The chromaffin cells in microcapsule survived well and could secrete dopamine.The HPLC results showed that the dopamine concentration arrived at the peak on 8th day of culture.Conclusion The dopamine secreted by microcapsulated bovine chromaffin cells can permeate APA membrane,and microcapsulated bovine chromaffin cell is a valuable source which may be used for cell therapy.

关 键 词：牛嗜铬细胞 微囊 多巴胺 

分 类 号：Q2-33[生物学—细胞生物学]