微囊技术对人嗜铬细胞和小鼠黑色素瘤细胞凋亡的影响

Effect of microencapsulation technique in the apoptosis of human chromaffin cells and mouse melanoma cells

目的:观察海藻酸钠-聚赖氨酸-海藻酸钠(APA)微囊对囊内细胞的存活、生长及凋亡的影响,以验证免疫隔离APA微胶囊的生物膜性和生物活性。方法:实验于2004-01/05在华中科技大学同济医学院附属同济医院麻醉科实验中心完成。取人嗜铬细胞原代培养,小鼠黑色素瘤细胞和小鼠成纤维细胞传代培养,将APA微囊分别包裹上述3种细胞并进行培养;根据3种细胞微囊化与否分成6组,利用MTT比色法检测各组细胞存活和生长情况,共观察9d,并用流式细胞仪检测各组细胞凋亡情况。结果:①在实验观察的9d内,人嗜铬细胞、小鼠黑色素瘤细胞和小鼠成纤维细胞可在微囊内以细胞团的形式生长、存活。②MTT比色法检测显示各组囊内细胞与相应组裸细胞的生长、存活情况比较差异无显著性意义(P>0.05)。③流式细胞仪检测各组囊内细胞与相应组裸细胞的凋亡百分率比较差异亦无显著性意义(P>0.05)。结论:制备的APA微囊化材料及制备方法对细胞活性、生长状况及凋亡情况无不良影响,表明包裹细胞的APA微囊有良好的生物相容性和生物活性。

AIM： To observe the survival, growth and apoptosis of alginate-polylysine-alginate （APA） microencapsulated cells, and verify the biological membrane and activity of APA microcapsules with immunoisolation. METHODS： The experiment was carried out in the test center, Department of Anesthesiology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology from January to May in 2004. Human chromaffin cells were harvested for primary culture, mouse melanoma cell and mouse fibroblast were subcultured, then three kinds of cells were encapsulated and cultured in APA microcapsules. According APA microencapsulation or not, three kinds of cells were divided into 6 groups and their survival and growth were measured for 9 days by MTT colorimetric assay. Flow cytometry was used to detect the cell apoptosis. RESULTS： 1.During 9-day observation, human chromaffin cells, mouse melanoma cell and mouse fibroblast in the microcapsules were all survived and grew in the clusters.2.MTT colorimetric assay showed that the differences of cell growth and survival were not significant between the microencapsulated group with APA and non-microencapsulated group with APA （P 〉 0.05）.3.Flow cytometry revealed that there was no significant difference in the rate of apoptosis between the microencapsulated group and non-microencapsulated group （P 〉 0.05）. CONCLUSION： APA microencapsulation technique has no harm to the activity, growth and apoptosis of cells, indicating good biological compatibility and activity of APA.