Department of Biochemical Engineering, University College London, Gordon Street, WC1H 0AH, London, United Kingdom.
Department of Nanobiomedical Science and BK21 Plus NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan 31116, Republic of Korea.
Biotechnol J. 2018 Feb;13(2). doi: 10.1002/biot.201700085. Epub 2018 Jan 25.
Human mesenchymal stromal cells (hMSCs) are excellent candidates for cell therapy but their expansion to desired clinical quantities can be compromised by ex vivo processing, due to differences between donor material and process variation. The aim of this article is to characterize growth kinetics of healthy baseline "reference" hMSCs using typical manual processing. Bone-marrow derived hMSCs from ten donors are isolated based on plastic adherence, expanded, and analyzed for their growth kinetics until passage 4. Results indicate that hMSC density decreases with overall time in culture (p < 0.001) but no significant differences are observed between successive passages after passage 1. In addition, fold increase in cell number dropped between passage 1 and 2 for three batches, which correlated to lower performance in total fold increase and expansion potential of these batches, suggesting that proliferative ability of hMSCs can be predicted at an early stage. An indicative bounded operating window is determined between passage 1 and 3 (PDL < 10), despite the high inter-donor variability present under standardized hMSC expansion conditions used. hMSC growth profile analysis will be of benefit to cell therapy manufacturing as a tool to predict culture performance and attainment of clinically-relevant yields, therefore stratifying the patient population based on early observation.
人源间充质基质细胞（hMSCs）是细胞治疗的优秀候选者，但由于供体材料的差异和体外处理过程中的变异性，其体外扩增到所需的临床数量可能会受到影响。本文的目的是使用典型的手动处理方法，对健康基线“参考”hMSCs 的生长动力学进行特征描述。从十个供体中分离出基于塑料贴附的骨髓源性 hMSCs，进行扩增，并分析其生长动力学，直到第 4 代。结果表明，hMSC 密度随培养时间的整体增加而降低（p<0.001），但在第 1 代之后的连续传代中没有观察到显著差异。此外，有三个批次的细胞数量在第 1 代和第 2 代之间的倍增减少，这与这些批次的总倍增和扩增潜力的性能较低相关，表明 hMSCs 的增殖能力可以在早期阶段进行预测。尽管在使用标准化的 hMSC 扩增条件下存在高供体间变异性，但在第 1 代和第 3 代（PDL<10）之间确定了一个有指示意义的有界操作窗口。hMSC 生长曲线分析将作为一种预测培养性能和达到临床相关产量的工具，对细胞治疗制造有益，从而根据早期观察结果对患者人群进行分层。
