成骨细胞在旋转壁式生物反应器内的大规模扩增  

Large Scale Expansion of Rat's Osteoblasts in Rotating Wall Vessel(RWV) Bioreactor

作　　者：宋克东[1] 刘天庆[1] 崔占峰[2] 李香琴[1] 葛丹[1] 孙相玉[1] 马学虎[1] 

机构地区：[1]大连理工大学干细胞与组织工程研究室,大连116024 [2]牛津大学工程科学系

出　　处：《中国生物医学工程学报》2005年第2期134-139,共6页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金两个基地国际合作项目 (2 0 0 2 0 0 8) ;辽宁省科学技术基金资助项目 (2 0 0 2 2 14 0 )。

摘　　要：目的　进行SD大鼠成骨细胞在旋转壁式生物反应器内大规模扩增培养的研究。方法　利用微载体悬浮培养法在旋转壁式生物反应器内大规模扩增成骨细胞 ,检测其组织形态和生物功能后。作为接种到支架材料上并于反应器内三维环境中培养组织工程骨的种子细胞。结果　成骨细胞在生物反应器中每代可以扩增十倍以上 ,同时经过倒置相差显微镜、SEM(扫描电镜 )以及ALP(碱性磷酸酶 )和MTT等生物学性能检测后 ,发现在旋转壁式生物反应器中三维培养的成骨细胞各种生物指标性能良好。结论　新型旋转壁式生物反应器可以提供低剪切力的培养环境 ,而且细胞之间有三维联系的机会 ,成骨细胞表现出良好的体外扩增能力 ,适于建立一种理想的成骨细胞体外扩增的三维培养体系。Purpose: Large scale expansion of rat's osteoblasts in rotating wall vessel (RWV) bioreactor was studied. Methods: Expand osteoblasts in large scale in RWV by using microcarrier in suspension culture. After evaluated histomorphology and biological functions, the expanded cells were seeded in the scaffolds and cultivated in the bioreactor. Results: Osteoblasts cultivated in the bioreactor in three dimensions were expanded more than ten times in one generation. The expanded cells performed well in major functions examined by inverted microscope in phase contrast observation, scanning electron microscope (SEM), alkaline phosphatase (ALP) test and MTT test. Conclusion: This novel RWV bioreactor provided the culture environment with relatively low shear force and good communication among cells in three-dimension, therefore osteoblasts showed a strong ability to be expanded in vitro. The RWV bioreactor showed great potential of being taken as an ideal three-dimensional culture system for osteoblasts to be expanded in vitro.

关 键 词：成骨细胞 微载体 体外培养 旋转壁式生物反应器 

分 类 号：R318[医药卫生—生物医学工程] Q813.11[医药卫生—基础医学]