化工学报 ›› 2022, Vol. 73 ›› Issue (4): 1546-1556.doi: 10.11949/0438-1157.20211574

• 流体力学与传递现象 • 上一篇    下一篇

摇床T25细胞培养瓶流体力学与传质特性研究

刘宏斐1(),李雪良1,钱钧弢2,刘金2,堵国成1,陈坚1()   

  1. 1.江南大学未来食品科学中心，江苏 无锡 214122
    2.迪必尔生物工程（上海）有限公司，上海 201611
  • 收稿日期:2021-11-09 修回日期:2021-12-21 出版日期:2022-04-05 发布日期:2022-04-25
  • 通讯作者: 陈坚 E-mail:18454300236@163.com;jchen@jiangnan.edu.cn
  • 作者简介:刘宏斐（1996—），女，硕士研究生，18454300236@163.com
  • 基金资助:
    江苏省自然科学基金前沿引领技术基础研究专项(BK20202002);中央高校基本科研业务费专项资金(JUSRP12011)

Hydrodynamics and mass transfer in rocking T25 cell culture flasks

Hongfei LIU1(),Xueliang LI1,Juntao QIAN2,Jin LIU2,Guocheng DU1,Jian CHEN1()   

  1. 1.Science Center for Future Foods, Jiangnan University, Wuxi 214122, Jiangsu, China
    2.T&J Bioengineering (Shanghai), Ltd. , Shanghai 201611, China
  • Received:2021-11-09 Revised:2021-12-21 Published:2022-04-05 Online:2022-04-25
  • Contact: Jian CHEN E-mail:18454300236@163.com;jchen@jiangnan.edu.cn

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摘要：

细胞培养技术是生物医药产业的支柱，以实现微小体积内的高密度、高通量细胞培养为目的，系统地研究了常用于单层静态培养的T25方瓶置于翘板摇床上在不同操作条件下的流体力学特性和传质性能。结果表明，振荡可以显著提高方瓶的传质速率并降低混合时间，使高密度培养成为可能，但瓶盖上的空气滤膜在高转速时成为传质速率的限制因素；培养瓶对称轴与摇床旋转轴平行时，其相对位置对混合和传质无明显影响，但当二者成45°角时，相同转速下混合时间显著缩短；使用自定义函数实现了基于动态网格的CFD模拟，对不同转速下方瓶内剪切应力和能量耗散在时间与空间上分布进行了分析，为基于T25培养瓶开发一次性高通量微型反应器提供了数据支持和理论基础。

关键词: T25方瓶, 细胞培养, 混合, 传质, 计算流体力学

Abstract:

Cell culture technology is the pillar of the biopharmaceutical industry. T-flasks as single-use consumables are typically used in single-layer static cultivation which produces limited cell density. In order to achieve high-throughput and high density cell culture in T-flasks, this study systematically investigates the hydrodynamics and mass transfer characteristics of a T25 flask placed on a rocking platform under different operating conditions. The results show that shaking can significantly increase the mass transfer rate of the square bottle and reduce the mixing time, making high-density culture possible. But the air filter on the bottle cap becomes the limiting factor for the mass transfer rate at high rotation speeds. The horizontal and vertical distances of the flask relative to the rocking axis had negligible effects on mass transfer and mixing, but placing the flask at a 45° angle and effectively using the walls of the flask as baffles, drastically reduces the mixing time at the same rocking speed. A user-defined function is used to implement a dynamic mesh to simulate the rocking movement of the T-flask in a CFD model. The CFD model enables the analysis of the shear stress and energy dissipation rates in the flask under different rocking speeds. This study provides the data and theoretical support for the further development of a T-flask-based high-throughput, single-use microbioreactor systems.

Key words: T25 flask, cell culture, mixing, mass transfer, CFD

中图分类号: 

  • TQ 021.1

图1

T25细胞培养瓶冷模实验装置"

图2

T25细胞培养瓶几何模型及网格划分"

图3

不同振荡转速与换气方式组合下T25培养瓶中溶氧变化a—置换氮气,开口无瓶盖;b—置换氮气,刺穿瓶盖通空气;c—置换氮气,盖瓶盖,不通空气;d—未置换氮气,刺穿瓶盖通空气"

图4

振荡转速对T25培养瓶表观传质系数的影响a—置换氮气,开口无瓶盖;b—置换氮气,刺穿瓶盖通空气;c—置换氮气,盖瓶盖,不通空气;d—未置换氮气,刺穿瓶盖通空气"

图5

70 r/min 时T25培养瓶混合情况及红色光吸收情况与CFD模拟结果的比较左—原始视频帧;中—红色光吸光度;右—CFD模拟结果"

图6

70 r/min时T25培养瓶混合过程红色光吸光度标准差"

图7

振荡转速和旋转轴位置对T25培养瓶混合情况的影响a—正放;b—培养瓶中心轴与摆动转轴成45°;c—垫高;d—偏左"

图8

玻璃珠对T25培养瓶混合时间的影响a—中央正放不加玻璃珠;b—中央正放加玻璃珠"

图9

80 r/min下不同网格数量下计算域内平均液速随时间变化曲线"

图10

不同模型与转速的CFD模拟结果"

图11

混合过程中培养瓶内不同位置示踪剂质量分数变化与红色吸收光标准差"

图12

T25培养瓶在20~80 r/min转速下气液界面状态"

图13

不同转速下T25培养瓶速度分布云图"

图14

不同转速下气液界面面积随时间的变化"

图15

转速对质量加权平均剪切应力影响"

图16

不同转速下T25培养瓶内剪切应力分布云图"

图17

转速对质量加权平均能量耗散率影响"

图18

T25培养瓶能量耗散率分布云图"

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