中空纤维膜在旋转壁式生物反应器内作为细胞载体的性能  

Cell Carrier Function of Hollow-fiber Membrane in Rotating Wall Vessel Bioreactor

作　　者：宋克东[1] 刘天庆[1] 赵虎[1] 李香琴[1] 崔占峰[2] 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,辽宁大连116024 [2]牛津大学工程科学系

出　　处：《高校化学工程学报》2007年第3期488-493,共6页Journal of Chemical Engineering of Chinese Universities

基　　金：国家自然科学基金(2002008);国家自然科学基金(30670525);校青年教师培养基金(893228)资助项目

摘　　要：利用中空纤维膜作为成骨细胞的载体,在旋转壁式中空纤维膜生物反应器(Rotating wall hollow-fiber membrane bioreactor,RWHMB)内对其进行大规模扩增,并在静态环境使用中空纤维膜和在旋转壁式生物反应器(Rotating wall vessel bioreactor,RWVB)中使用微载体以相同条件对照培养。每隔12h对细胞进行取样,培养5d后对细胞进行收获。对所扩增的细胞分别进行扩增能力、扫描电镜(SEM)观察、苏木精-伊红染色、碱性磷酸酶(ALP)染色、钙化结节染色等生物学性能检测。结果显示在RWHMB内动态培养的成骨细胞生长良好,分泌大量的细胞外基质。证实RWHMB能结合RWVB和中空纤维膜的优点,较好地模拟体内环境,对所培养的细胞施加适当的流体应力刺激,促进细胞的新陈代谢和物质交换,更有利于细胞的生长及增殖。Large-scale expansion of SD （Sprague-Dawley） rat＇s osteoblasts was studied in rotating wall hollow-fiber membrane bioreactor （RWHMB） by using hollow-fiber membrane as the carrier. For the sake of contrast, cells were also expanded in T-flask using hollow-fiber membrane as carrier and in rotating wall vessel bioreactor （RWVB） with microcarrier as carrier. During the culture period, the cells were sampled every 12 hr, and after 5 days, the cells were harvested and evaluated with scanning electron microscope （SEM）, hematoxylin-eosin （HE） staining and alkaline phosphatase （ALP） staining. Moreover, von-Kossa staining and Alizarin Red S staining were carried out for mineralized nodules formation. The results show that in RWHMB, the cells present better morphology and vitality and secrete much more extracellular matrix. It was concluded that the RWHMB combines the advantages of rotating wall vessel and hollow-fiber membrane bioreactors. The hydrodynamic stimulation within it accelerates the metabolism of osteoblast and mass transfer, which is propitious to the cell differentiation and proliferation.

关 键 词：中空纤维膜 微载体 生物反应器 成骨细胞 扩增 

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