皮肤成纤维细胞在生物反应器中的生长和扩增  

GROWTH AND EXPANSION OF HUMAN DERMAL FIBROBLASTS IN A BIOREACTOR

作　　者：周燕[1] 邓明安[1] 华平[1] 谭文松[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《中国修复重建外科杂志》2003年第2期89-92,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家高技术研究发展计划课题 (2 0 0 1 AA2 1 0 641 ) ;国家重点基础研究发展规划项目 (973) :组织工程的基本科学问题(G1 9990 5430 9)~~

摘　　要：目的　探讨皮肤成纤维细胞在生物反应器中批式和间歇换液培养过程的生长、扩增与代谢特性。方法　将幼儿包皮组织中分离皮肤成纤维细胞接种到含 5 mg/ ml微载体浓度的 1.5升 Celli Gen生物反应器中 ,测定在间歇换液培养和批式培养过程中细胞生长密度、葡萄糖消耗及乳酸生成 ,同时与方瓶和转瓶中间歇换液培养结果进行比较。　结果　在生物反应器间歇换液培养过程中 ,成纤维细胞生长的最终密度达到 2 .0 8× 10 6 / ml,扩增 2 9.7倍 ,是批式培养的 1.81倍。与间歇换液的方瓶和转瓶培养结果相比 ,由于消除了营养和贴壁表面的限制 ,细胞密度分别提高9.16倍和 1.4 3倍。　结论　利用生物反应器和微载体悬浮培养人皮肤成纤维细胞 ,是大量制备组织工程种子细胞的一种有效方法。Objective To investigate the growth, expansion, and metabolic characteristics of the human dermal fibroblasts cultured in a bioreactor with batch and medium exchange modes. Methods Human dermal fibroblasts separated from foreskin were seeded into a 1.5 liter CelliGen bioreactor with 5mg/ml of microcarriers. The cell growth, glucose consumption and lactate accumulation in both batch and medium exchange cultures were measured. Results The growth density of fibroblasts cultured in the bioreactor with medium exchange mode reached 2.08×10 6 cell/ml, expanded 29.7 folds, which was 1.81 times as high as that in batch culture. By comparison with the results obtained in T flasks and spinners under the same medium exchange conditions, the cell density in the bioreactor was 9.16 and 1.43 times as high as those in T flasks and spinners respectively owing to that the limitation effect of the attachment surface, nutrient exhaust, and by product accumulation on the growth of fibroblasts in the bioreactor by using microcarriers, medium exchange, as well as gas aeration was elimnated. Conclusion The above results indicate that suspended cultures with microcarriers in bioreactors are an effective approach to provide large amounts of seeding cells for tissue engineering.

关 键 词：皮肤成纤维细胞 生物反应器 细胞生长 细胞扩增 细胞代谢 皮肤移植 

分 类 号：R318.19[医药卫生—生物医学工程]