Hupfeld Julia, Gorr Ingo H, Schwald Christian, Beaucamp Nicola, Wiechmann Kornelius, Kuentzer Karin, Huss Ralf, Rieger Bernhard, Neubauer Markus, Wegmeyer Heike
Pharma Research and Early Development (pRED), Roche Diagnostics GmbH, Nonnenwald 2, Penzberg, 82377, Germany.
Biotechnol Bioeng. 2014 Nov;111(11):2290-302. doi: 10.1002/bit.25281. Epub 2014 Jul 14.
Mesenchymal stromal cells (MSCs) are promising candidates for cell therapy. Their therapeutic use requires extensive expansion to obtain a sufficiently high number of cells for clinical applications. State-of-the-art expansion systems, that is, primarily culture flask-based systems, are limited regarding scale-up, automation, and reproducibility. To overcome this bottleneck, microcarrier (MC)-based expansion processes have been developed. For the first time, MSCs from the perinatal sources umbilical cord (UC) and amniotic membrane (AM) were expanded on MCs. This study focuses on the comparison of flask- and Cytodex 1 MC-expanded MSCs by evaluating the influence of the expansion process on biological MSC characteristics. Furthermore, we tested the hypothesis to obtain more homogeneous MSC preparations by expanding cells on MCs in controlled large-scale bioreactors. MSCs were extensively characterized determining morphology, cell growth, surface marker expression, and functional properties such as differentiation capacity, secretion of paracrine factors, and gene expression. Based on their gene expression profile MSCs from different donors and sources clearly clustered in distinct groups solely depending on the expansion process-MC or flask culture. MC- and flask-expanded MSCs significantly differed from each other regarding surface markers and both paracrine factors and gene expression profiles. Furthermore, based on gene expression analysis, MC cultivation of MSCs in controlled bioreactor systems resulted in less heterogeneity between cells from different donors. In conclusion, MC-based MSC expansion in controlled bioreactors has the potential to reliably produce MSCs with altered characteristics and functions as compared to flask-expanded MSCs. These findings may be useful for the generation of MSCs with tailored properties for clinical applications.
间充质基质细胞（MSCs）是细胞治疗中很有前景的候选细胞。其治疗用途需要大量扩增以获得足够数量的细胞用于临床应用。目前最先进的扩增系统，即主要基于培养瓶的系统，在扩大规模、自动化和可重复性方面存在局限性。为了克服这一瓶颈，已开发出基于微载体（MC）的扩增工艺。首次在微载体上扩增了来自围产期来源脐带（UC）和羊膜（AM）的间充质基质细胞。本研究通过评估扩增过程对间充质基质细胞生物学特性的影响，重点比较了在培养瓶和Cytodex 1微载体上扩增的间充质基质细胞。此外，我们测试了通过在受控的大规模生物反应器中的微载体上扩增细胞来获得更均匀的间充质基质细胞制剂的假设。对间充质基质细胞进行了广泛的表征，包括确定形态、细胞生长、表面标志物表达以及功能特性，如分化能力、旁分泌因子分泌和基因表达。基于它们的基因表达谱，来自不同供体和来源的间充质基质细胞仅根据扩增过程（微载体或培养瓶培养）明显聚集在不同的组中。微载体扩增和培养瓶扩增的间充质基质细胞在表面标志物、旁分泌因子和基因表达谱方面存在显著差异。此外，基于基因表达分析，在受控生物反应器系统中对间充质基质细胞进行微载体培养导致不同供体细胞之间的异质性降低。总之，与培养瓶扩增的间充质基质细胞相比，在受控生物反应器中基于微载体的间充质基质细胞扩增有可能可靠地产生具有改变的特性和功能的间充质基质细胞。这些发现可能有助于生成具有适合临床应用特性的间充质基质细胞。
