微囊化牛嗜铬细胞及冷冻保存的体外培养研究  

Study of microencavsulated bovine ehromaffin cells and its cryopreservation in vitro

作　　者：杨学伟[1] 黄乔东[2] 章乐虹[1] 胡以则[1] 

机构地区：[1]广州医学院第二附属医院普通外科,510260 [2]广州医学院第二附属医疼痛科院,510260

出　　处：《国际医药卫生导报》2011年第14期1668-1670,共3页International Medicine and Health Guidance News

基　　金：广州市医药卫生科技项目（2007-YB-165）

摘　　要：目的 观察体外培养的微囊化牛嗜铬细胞及其冷冻保存复苏后的细胞活力,探索微囊化牛嗜铬细胞的冻存保存方法,为将来进行大规模细胞移植治疗疼痛奠定临床基础.方法 经胶原酶消化成单细胞,微囊包裹,并取部分进行冷冻保存及复温,观察细胞的生长情况;放免法分别检测其培养液中去甲肾上腺素、甲-脑啡呔的含量和在高钾、乙酰胆碱刺激下的分泌量来观察其功能活性.结果 微囊化牛嗜铬细胞及其冷冻复苏后的细胞形态结构完整,生长良好;冻存复苏后的微囊保留了儿茶酚胺和甲-脑啡呔的的分泌功能,基础与刺激分泌量大约为冻存前微囊化牛嗜铬细胞分泌量的92%;同时刺激后去甲肾上腺素、甲-脑啡呔水平较其基础值均有明显增加（P＜0.01）.结论 微囊化牛嗜铬细胞及其冷冻复苏后的细胞具有良好的细胞功能,冷冻保存的微囊化牛嗜铬细胞是有效和成功的.Objective To observe the viability of microencapsulated bovine chromaffin cells and their viability after cryopreservation, explore a cryopreservation mean for the microencapsulated chomaffin cells, and build a clinical foundation for the future large-scale clinical cell transplantation for treatment of pain. Methods Bovine adrenal medullary was digested into single chromaffin with collagenase digestion, then microencapsulated it. The cells were cryopreservated and some of them were thawed for in vitro culture to observe the cell growth; Culture medium was collected for the analysis of norepinephrine and methyl-endomorphin concentration by radioimmunoassay. Cell viability was evaluated by the secretion under high potassium and acetylcholine stimulation. Results Microencapsulated bovine chromaffin cells displayed their morphology structural integrity and functioned well after cryopreservation. The cells maintained the ability to secrete catecholamines and methyl-endomorphin. The basic and stimulated secretion volume of microencapsulated bovine chromaffin cells recovered from cryopreservation were 92% of that of fresh chromaffin cells. The norepinephrine and methyl-endomorphin levels were significantly higher after stimulation （P〈0.01）. Conclusion Microencapsulated bovine chromaffin cells and thawed cells from cryopreservation show the ability to secret both catecholamines and methyl-endomorphin well, which demonstrated that the microencapsulation and cryopreservation are useful method to preserve bovine chromaffin cells.

关 键 词：海藻酸钠-多聚赖氨酸-海藻酸钠微囊 牛嗜铬细胞 冻存 

分 类 号：R4[医药卫生—临床医学]