A mechanistic model of erythroblast growth inhibition providing a framework for optimisation of cell therapy manufacturing
A mechanistic model of erythroblast growth inhibition providing a framework for optimisation of cell therapy manufacturing
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DOI：
10.1016/j.bej.2018.01.033
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发表时间：
2018-05-15
影响因子：
3.9
通讯作者：
Thomas, Robert J.
中科院分区：
文献类型：
--
作者：
Glen, Katie E.;Cheeseman, Elizabeth A.;Thomas, Robert J.
在体外制造红细胞为基础的产品需要高效的红母细胞培养经济可行性。以前的研究表明，可扩展生物反应器中红母细胞培养的效率主要不受传质、培养基成分的可用性或公认的抑制性代谢物或细胞因子的限制。我们开发了一个动态机制模型，描述了一个自分泌反馈回路，其中细胞衍生因子在培养基中积累，导致可逆的红母细胞生长抑制。细胞表现出两个生长阶段：一个相对不受抑制的阶段，接着是一个受抑制的阶段。抑制期间的细胞周期分析发现，细胞在S期有轻微的积累，这与生长因子或营养剥夺所预期的G1期积累不同。供体对供体生长速率的显著差异(平均0.047 h（-1)，标准差0.008 h(-1)）要求对不同供体的生长速率参数进行调整。该模型可用于预测完全介质交换条件下的生长行为，但部分介质交换条件下的预测能力有所下降。该模型可以预测从早成熟到晚成熟的一系列表型成熟的红母细胞的生长拐点：然而，在较成熟的细胞中，生长的第二阶段有很大的不同，观察到的抑制较小。该模型提供了一个基于生产时间成本和投入消耗品成本的养殖经济优化框架。它还提供了一个框架来评估生物工艺工程在介质设计或细胞修改与操作优化方面的好处，这取决于工艺开发商的具体成本方案。(C) 2018作者。Elsevier B.V.出版
Manufacture of Red Blood Cell based products in vitro requires highly efficient erythroblast culture for economic viability. It has previously been shown that efficiency of erythroblast culture in scalable bioreactors is not primarily limited by mass transfer, availability of medium components, or commonly recognised inhibitory metabolites or cytokines. We have developed a dynamic mechanistic model that describes an autocrine feedback loop in which a cell-derived factor accumulates in culture medium resulting in reversible erythroblast growth inhibition. Cells exhibited two phases of growth: a relatively uninhibited followed by an inhibited phase. Cell cycle analysis during inhibition identified slight accumulation of cells in S phase, distinct from the G1 accumulation anticipated in growth factor or nutrient deprivation. Substantial donor to donor growth rate variability (mean 0.047 h(-1), standard deviation 0.008 h(-1)) required the growth rate parameter to be refitted for different donors. The model could then be used to predict growth behaviour with full medium exchange, but showed some reduced predictive ability after partial medium exchange. The model could predict the growth inflexion point over a range of phenotypic maturities from early to late maturity erythroblasts: however the secondary phase of growth differed substantially with less inhibition observed in more mature cells. The model provided a framework to optimise culture economics based on cost of production time and input consumables. It also provided a framework to evaluate the benefits of biological process engineering in medium design or cell modification vs. operational optimisation depending on the specific cost scenario of a process developer. (C) 2018 The Author(s). Published by Elsevier B.V.