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New miniaturized hollow-fiber bioreactor for in vivo like cell culture,cell expansion, and production of cell-derived products

机译:新型微型中空纤维生物反应器,用于体内细胞培养,细胞扩增和细胞衍生产品的生产

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We have developed a miniaturized hollow-fiber bioreactor system for mammalian cell culture with a volume of 1 mL. Cell and medium compartments of the bioreactor are separated by a semipermeable membrane, and oxygenation of the cell compartment is accomplished using an oxygenation membrane. As a result of the geometry of the transparent housing, cells can be observed by microscopy during culture. The leukemic cell lines CCRF-CEM, HL-60, and REH were cultivated up to densities of 3.5 x 10(7)/ mL without medium change or manipulation of the cells. As shown using CCRF-CEM cells, growth in the bioreactor was strongly influenced and could be controlled by the medium flow rate. As a consequence, consumption of glucose and generation of lactate varied with flow rate. Depending on the molecular size cutoff of the membranes used, added growth factors such as GM-CSF, as well as factors secreted from the cells, are retained in the cell compartment for up to 1 week. This new miniaturized hollow-fiber bioreactor offers advantages in tissue engineering by continuous nutrient supply for cells in high density, retention of added or autocrine produced factors, and undisturbed long-term culture in a closed system.
机译:我们已经开发了用于哺乳动物细胞培养的微型中空纤维生物反应器系统,体积为1 mL。生物反应器的细胞室和培养基室被半透膜隔开,并且细胞室的氧合是使用氧合膜完成的。由于透明外壳的几何形状,可以在培养过程中通过显微镜观察细胞。将白血病细胞系CCRF-CEM,HL-60和REH培养至密度为3.5 x 10(7)/ mL,无需更换培养基或操作细胞。如使用CCRF-CEM细胞所显示的,生物反应器中的生长受到强烈影响,并且可以通过培养基流速来控制。结果,葡萄糖的消耗和乳酸的产生随流速而变化。根据所用膜的分子大小截断,添加的生长因子(例如GM-CSF)以及细胞分泌的因子会在细胞室内保留长达1周。这种新的小型化中空纤维生物反应器通过为细胞提供高密度的持续营养,保留添加的或自分泌的生产因子以及在封闭系统中不受干扰的长期培养,为组织工程提供了优势。

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