Song Kedong, Yan Xinyu, Zhang Yu, Song Fei, Lim Mayasari, Fang Meiyun, Shi Fangxin, Wang Ling, Liu Tianqing
State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian, 116024, China,
Bioprocess Biosyst Eng. 2015 Aug;38(8):1527-40. doi: 10.1007/s00449-015-1395-6. Epub 2015 Apr 14.
Currently, RWVB (Rotating wall vessel bioreactor) combined with a microcarrier used for in vitro expansion revealed that the suspended cells attached on the microcarrier will collide with outer and inner cylinders of RWVB inevitably, which leads to harmful results to the cells. Considering this, hollow fiber (HF) membrane module treated as a cell carrier is adopted to combine with RWVB to form a novel rotating wall hollow fiber membrane bioreactor (RWHMB) to avoid aforementioned harmful collision, since the cells cultured inside this bioreactor will mainly adhere to large specific surface of hollow fiber membrane module. Prior to cell experiment, mathematical simulations concerned with flow field inside RWHMB are performed by CFD, which includes the distributions of the total pressure, velocity, and shear stress with the variation of rotating speeds and directions, as well as the radial location and diameter of hollow fiber membrane. To further confirm the feasible parameters getting from the simulation, this RWHMB is adopted to expand osteoblasts isolated from SD rats within its dynamic conditions. Cell expansion in T-flask is carried out as a negative control. The results showed that with the same rotating direction and speed of 10 rpm, inner and outer cylinders of RWHMB generated cyclical stress stimulus, which was acceptable to cell expansion and facilitated the secretion of extracellular matrix. Besides, hollow fiber membrane carrier with a diameter of 0.2 mm has an excellent biocompatibility and their radial locations presented a tiny influence on flow field inside the culture chamber.
目前,旋转壁式生物反应器(RWVB)与用于体外扩增的微载体相结合的研究表明,附着在微载体上的悬浮细胞不可避免地会与RWVB的内筒和外筒发生碰撞,这会对细胞产生有害影响。考虑到这一点,采用中空纤维(HF)膜组件作为细胞载体与RWVB相结合,形成一种新型的旋转壁中空纤维膜生物反应器(RWHMB),以避免上述有害碰撞,因为在该生物反应器内培养的细胞将主要附着在中空纤维膜组件的大比表面积上。在进行细胞实验之前,通过计算流体力学(CFD)对RWHMB内部的流场进行了数学模拟,其中包括总压力、速度和剪切应力随转速和方向变化的分布,以及中空纤维膜的径向位置和直径。为了进一步确认从模拟中获得的可行参数,采用该RWHMB在其动态条件下扩增从SD大鼠分离的成骨细胞。在T型烧瓶中进行细胞扩增作为阴性对照。结果表明,在相同的旋转方向和10 rpm的转速下,RWHMB的内筒和外筒产生周期性应力刺激,这对细胞扩增是可接受的,并促进了细胞外基质的分泌。此外,直径为0.2 mm的中空纤维膜载体具有优异的生物相容性,其径向位置对培养室内的流场影响很小。