微囊化人肾上腺嗜铬细胞瘤细胞在人工脑脊液中的生长和分泌特征  

Growth and secretion features of microencapsulated human adrenal pheochromocytoma cells in artificial cerebrospinal fluid

作　　者：栗晓[1] 张新胜[1] 罗海杰[1] 栾烁[1] 万青[1] 伍少玲[1] 马超[1] 

机构地区：[1]中山大学孙逸仙纪念医院康复医学科,广东省广州市510120

出　　处：《中国组织工程研究》2015年第38期6114-6120,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81171469);广东省医学科学技术研究基金项目(A2015308)~~

摘　　要：背景:微囊化细胞是目前常用的免疫隔离工具,可以解决细胞蛛网膜下腔移植存在的免疫排斥问题,然而微囊化对人嗜铬细胞瘤细胞分泌功能的影响尚不明确。目的:观察微囊化的人嗜铬细胞瘤细胞在人工脑脊液中的生长及分泌功能。方法:取手术切除的人嗜铬细胞瘤组织,采用连续分次胶原酶消化法分离人嗜铬细胞瘤细胞,并用人工脑脊液进行原代细胞培养。用海藻酸盐-聚赖氨酸-海藻酸盐微囊包裹原代培养的人嗜铬细胞瘤细胞,用倒置相差显微镜观察人工脑脊液中的微囊化和非微囊化的人嗜铬细胞瘤细胞的形态;用CCK-8试剂盒测定人嗜铬细胞瘤细胞的增殖情况;用Elisa试剂盒检测微囊化和非微囊化的人嗜铬细胞瘤细胞分泌的去甲肾上腺素和甲硫氨酸脑啡肽浓度。结果与结论:微囊化的人嗜铬细胞瘤细胞镜下呈悬浮、聚集生长状态,细胞突起观察不明显。与非微囊化组相比,微囊化人嗜铬细胞瘤细胞增殖较快,甲硫氨酸脑啡肽和去甲肾上腺素的分泌量较多。但不同病例来源的细胞,甲硫氨酸脑啡肽和去甲肾上腺素的分泌量差异较大。提示微囊化的人嗜铬细胞瘤细胞在人工脑脊液中具有良好的生长和分泌功能,并且不同病例来源的人嗜铬细胞瘤细胞可以相对稳定地分泌甲硫氨酸脑啡肽和去甲肾上腺素。BACKGROUND： Microencapsulated cells are commonly used as a tool to overcome immune rejection after subarachnoid transplantation. However, the effect of microencapsulation on the secretion of human pheochromocytoma cells is unclear. OBJECTIVE： To observe the growth and secretion of primarily microencapsulated cultured human pheochromocytoma cells in artificial cerebrospinal fluid. METHODS： The human pheochromocytoma tissues were digested successively to isolate human pheochromocytoma cells that were then cultured in artificial cerebrospinal fluid. Primary cells were covered with alginate-polylysine-alginate microcapsules, and then the cell morphology was observed with inverted phase contrast microscope. Levels of met-enkephalin and norepinephrine in cell culture medium were detected by enzyme-labeled immunosorbent assay（ELISA）. We used cell counting kit-8 colorimetric assay to obtain the growth curve of human pheochromocytoma cells in artificial cerebrospinal fluid. RESULTS AND CONCLUSION： Microcapsulated human pheochromocytoma cells were in suspension and the process outgrowth increased slowly. Compared with non-microcapsulated cells, the proliferation rate of microcapsulated cells increased significantly. ELISA results revealed a significant increase in the levels of met-enkephalin and norepinephrine secreted from the microencapsulated cells compared to thenon-microcapsule group. There was a wide variation in contents of met-enkephalin and norepinephrine from different tumors. These findings indicate that microencapsulated human pheochromocytoma cells can survive well and have good secretion function in artificial cerebrospinal fluid, and human pheochromocytoma cells from different tumor tissues have stable secretory function.

关 键 词：PC12细胞 脑啡肽 甲硫氨酸 去甲肾上腺素 生物材料 材料相容性 人嗜铬细胞瘤细胞 原代培养 人工脑脊液 微胶囊 微囊化细胞 免疫隔离 甲硫氨酸脑啡肽 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]