Synchronized mammalian cell culture: Part I—A physical strategy for synchronized cultivation under physiological conditions
Synchronized mammalian cell culture: Part I—A physical strategy for synchronized cultivation under physiological conditions
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同步哺乳动物细胞培养：第 IâA 部分生理条件下同步培养的物理策略
DOI：
10.1002/btpr.1944
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发表时间：
2015
影响因子：
2.9
通讯作者：
Pörtner
中科院分区：
文献类型：
--
作者：
Platas Barradas;Becker;Bahnemann;Pörtner
哺乳动物细胞培养过程的常规分析和优化程序大多将培养物视为同质群体。因此，重点是细胞生理学和代谢，细胞系开发和过程控制策略。然而，不同亚群之间细胞特性的潜在变化对培养的影响尚未得到系统评价。形成这种亚群的一个主要原因是所有细胞在细胞周期中的进展。细胞行为中潜在细胞周期特异性变化与大规模工艺条件的相互作用可以通过（部分）同步培养以及随后的群体分辨模型分析来最佳确定。因此，希望以最小的扰动同步培养物，这可以使用物理选择方法获得不同的产量和质量，但不能使用常用的化学或全培养方法。传统的非同步方法与随后的细胞特异性，例如，流式细胞术分析，只能解决细胞周期的细胞限制效应。在这项工作中，我们证明逆流离心淘洗是一种有用的物理方法，可以在细胞周期的不同阶段富集哺乳动物细胞群，这些细胞群可以在生理条件下在生物反应器中进一步培养以同步生长。所提出的组合方法与其他物理选择方法形成对比，特别是在可实现的产量方面，这使得它适合于生物反应器规模的培养。如用两种工业细胞系（CH0-K1和人AGE 1.HN）所示，可以获得同步接种物，其中G1期的总体同步度高达82%，S期为53%，而G2期为60%。 富集因子分别为1.71、1.79和4.24。细胞能够在生物反应器中同步生长几个细胞周期。该策略结合随附论文中描述的群体分辨模型分析和参数提取，为在生理条件下在细胞周期和群体水平上研究细胞系和过程提供了新的可能性。© 2014美国化学工程师学会Biotechnol。程序：31：165 - 174，2015
Conventional analysis and optimization procedures of mammalian cell culture processes mostly treat the culture as a homogeneous population. Hence, the focus is on cell physiology and metabolism, cell line development, and process control strategy. Impact on cultivations caused by potential variations in cellular properties between different subpopulations, however, has not yet been evaluated systematically. One main cause for the formation of such subpopulations is the progress of all cells through the cell cycle. The interaction of potential cell cycle specific variations in the cell behavior with large‐scale process conditions can be optimally determined by means of (partially) synchronized cultivations, with subsequent population resolved model analysis. Therefore, it is desirable to synchronize a culture with minimal perturbation, which is possible with different yield and quality using physical selection methods, but not with frequently used chemical or whole‐culture methods. Conventional nonsynchronizing methods with subsequent cell‐specific, for example, flow cytometric analysis, can only resolve cell‐limited effects of the cell cycle. In this work, we demonstrate countercurrent‐flow centrifugal elutriation as a useful physical method to enrich mammalian cell populations within different phases of a cell cycle, which can be further cultivated for synchronized growth in bioreactors under physiological conditions. The presented combined approach contrasts with other physical selection methods especially with respect to the achievable yield, which makes it suitable for bioreactor scale cultivations. As shown with two industrial cell lines (CHO‐K1 and human AGE1.HN), synchronous inocula can be obtained with overall synchrony degrees of up to 82% in the G1 phase, 53% in the S phase and 60% in the phase, with enrichment factors ( ) of 1.71, 1.79, and 4.24 respectively. Cells are able to grow with synchrony in bioreactors over several cell cycles. This strategy, combined with population‐resolved model analysis and parameter extraction as described in the accompanying paper, offers new possibilities for studies of cell lines and processes at levels of cell cycle and population under physiological conditions. © 2014 American Institute of Chemical EngineersBiotechnol. Prog., 31:165–174, 2015
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影响因子：
--
作者：
Platas Barradas O;Jandt U;Hass R;Kasper C;Sandig V;Pörtner R;Zeng AP
通讯作者：
Zeng AP
标准化摇匀以同步培养的 CHO 细胞。
DOI：
--
发表时间：
1983
期刊：
Mutation research
影响因子：
--
作者：
T. Zwanenburg
通讯作者：
T. Zwanenburg
使用真核“婴儿机器”实现干扰最小、多周期且可重复的同步。
影响因子：
4
作者：
S. Cooper
通讯作者：
S. Cooper
影响因子：
11.2
作者：
K. Keyomarsi;L. Sandoval;V. Band;A. Pardee
通讯作者：
K. Keyomarsi;L. Sandoval;V. Band;A. Pardee
通过速度沉降技术同步小鼠 L 细胞。
影响因子：
3.4
作者：
H. Macdonald;R. Miller
通讯作者：
R. Miller