Monitoring the ex‐vivo expansion of human mesenchymal stem/stromal cells in xeno‐free microcarrier‐based reactor systems by MIR spectroscopy

Abstract

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 (RMSECV ≤ 4.7%, ≤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. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:447-455, 2016.

Mesenchymal stem cell cultivation in electrospun scaffolds: mechanistic modeling for tissue engineering

“…4a) have been widely used to expand MSCs in commercial microspheres, which are also called microcarriers, due to the enhanced mass transport inside the constructs and resultant higher cell growth [110]. In order to optimize MSC growth, different microcarriers [111], culture medium [111][112][113], and shear stress levels [114] have been studied in stirred bioreactors. However, contrary to microcarriers, tissue substitutes require appropriate geometry and functions, usually being cultivated in dynamic systems with perfusion and rotation (Fig.…”

A phenotypic screening bioassay forEscherichia colistress and antibiotic responses based on Fourier‐transform infrared (FTIR) spectroscopy and multivariate analysis

“…In that regard, Fourier‐transform infrared spectroscopy (FTIRS) acquires phenotypic profiles in a high‐throughput, rapid, label‐free, automatable, considerably inexpensive and reasonably simple mode (Sales et al ). FTIRS may be applied to discriminate and quantify diverse molecules, from proteins, nucleic acids, lipids through to diverse metabolites (Lopes et al ; Rosa et al ; Sampaio et al ), with the advantage of representing the whole omics of a cell (Bellisola and Sorio ). However, this implies a priori laborious calibration procedures for molecule identification and quantification.…”

The Challenge of Human Mesenchymal Stromal Cell Expansion: Current and Prospective Answers

“…Mid-infrared spectroscopy combined with multivariate data analysis is another promising online tool to optimize process monitoring for spinner cultures, particularly in the context of process prediction, contamination risks, speed and economic modeling. An optimized partial least squares regression model has been used to estimate glucose, lactate and ammonia concentrations [88].…”