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首页> 外文期刊>Biotechnology Progress >Monitoring the ex-vivo expansion of human mesenchymal stem/stromal cells in xeno-free microcarrier-based reactor systems by MIR spectroscopy
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Monitoring the ex-vivo expansion of human mesenchymal stem/stromal cells in xeno-free microcarrier-based reactor systems by MIR spectroscopy

机译：通过MIR光谱监测无异源微载体反应器系统中人间充质干/基质细胞的体外扩增

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Human mesenchymal stem/stromal cells (MSCs) have received considerable attention in the field of cell-based therapies due to their high differentiation potential and ability to modulate immune responses. However, since these cells can only be isolated in very low quantities, successful realization of these therapies requires MSCs ex-vivo expansion to achieve relevant cell doses. The metabolic activity is one of the parameters often monitored during MSCs cultivation by using expensive multi-analytical methods, some of them time-consuming. The present work evaluates the use of mid-infrared (MIR) spectroscopy, through rapid and economic high-throughput analyses associated to multivariate data analysis, to monitor three different MSCs cultivation runs conducted in spinner flasks, under xeno-free culture conditions, which differ in the type of microcarriers used and the culture feeding strategy applied. After evaluating diverse spectral preprocessing techniques, the optimized partial least square (PLS) regression models based on the MIR spectra to estimate the glucose, lactate and ammonia concentrations yielded high coefficients of determination (R(2)0.98, 0.98, and 0.94, respectively) and low prediction errors (RMSECV4.7%, 4.4% and 5.7%, respectively). Besides PLS models valid for specific expansion protocols, a robust model simultaneously valid for the three processes was also built for predicting glucose, lactate and ammonia, yielding a R-2 of 0.95, 0.97 and 0.86, and a RMSECV of 0.33, 0.57, and 0.09 mM, respectively. Therefore, MIR spectroscopy combined with multivariate data analysis represents a promising tool for both optimization and control of MSCs expansion processes. (c) 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:447-455, 2016
机译：人间充质干/基质细胞（MSC）由于其高分化潜力和调节免疫应答的能力，在基于细胞的治疗领域受到了广泛关注。但是，由于这些细胞只能以非常低的量分离出来，因此，成功实现这些疗法需要MSC进行离体扩增以获得相关的细胞剂量。代谢活性是在MSCs培养过程中经常通过使用昂贵的多分析方法监测的参数之一，其中一些方法很耗时。本工作通过与多变量数据分析相关的快速，经济的高通量分析，评估中红外光谱的使用，以监测在无异种培养条件下在旋转瓶中进行的三种不同的MSCs培养运行，这些条件不同所用微载体的类型以及所采用的培养物喂养策略。在评估了各种光谱预处理技术之后，基于MIR光谱的最佳偏最小二乘（PLS）回归模型可以估算葡萄糖，乳酸盐和氨的浓度，从而得出较高的测定系数（分别为R（2）0.98、0.98和0.94）。和较低的预测误差（分别为RMSECV4.7％，4.4％和5.7％）。除了适用于特定扩展协议的PLS模型外，还构建了同时适用于三个过程的健壮模型来预测葡萄糖，乳酸和氨，得出的R-2为0.95、0.97和0.86，RMSECV为0.33、0.57和分别为0.09 mM。因此，MIR光谱技术与多变量数据分析相结合代表了一种用于优化和控制MSC扩增过程的有前途的工具。 （c）2016美国化学工程师学会生物技术学会。 Prog。，32：447-455，2016

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