Characterization of human mesenchymal stem cells from multiple donors and the implications for large scale bioprocess development
Characterization of human mesenchymal stem cells from multiple donors and the implications for large scale bioprocess development
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DOI：
10.1016/j.bej.2015.06.018
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发表时间：
2016-04-15
影响因子：
3.9
通讯作者：
Hewitt, Christopher J.
中科院分区：
文献类型：
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作者：
Heathman, Thomas R. J.;Rafiq, Qasim A.;Hewitt, Christopher J.
基于细胞的疗法有可能为全球医疗保健做出贡献，因此活细胞和组织的使用可以用作药物疗法。尽管有这样的潜力，但在这一新兴领域的全部价值得以实现之前，仍然存在许多挑战。对来自多个捐赠者的细胞治疗生物过程的输入材料的特征是必要的，以确定和理解输入变化对过程开发的潜在影响。在这项工作中，我们对来自多个捐赠者的骨髓来源的人间充质干细胞(BM-hMSCs)进行了表征，并讨论了可测量的输入差异对开发基于自体和同种异体细胞的治疗制造过程的影响。5株BM-hMSC在30天内的累计群体倍增幅度为5.93，培养结束时集落形成率相差18.2%，产生IL-6的细胞株之间相差55.1%。已经证明，这种变异导致这些供体hMSC系之间的加工时间范围超过13天，当生产来自多个患者的产品时，会产生潜在的批次计时问题。所有BM-hMSC供体系均符合ISCT标准，但在细胞形态上存在差异。代谢产物分析显示，不同供体的hMSCs的葡萄糖消耗量为26.98pmocell(-1)天(-1)，乳酸生成量为29.45pmocell(-1)天(-1)，氨氮生成量为1.35pmocell(-1)天(-1)，说明供体在整个扩增过程中的变异程度。在工艺开发过程中测量信息丰富的产品属性将有助于朝着一致的制造工艺取得进展，这是基于细胞的翻译疗法的关键一步。(C)2015年提交人。爱思唯尔出版公司(Elsevier B.V.)
Cell-based therapies have the potential to contribute to global healthcare, whereby the use of living cells and tissues can be used as medicinal therapies. Despite this potential, many challenges remain before the full value of this emerging field can be realized. The characterization of input material-for cell-based therapy bioprocesses from multiple donors is necessary to identify and understand the potential implications of input variation on process development. In this work, we have characterized bone marrow derived human mesenchymal stem cells (BM-hMSCs) from multiple donors and discussed the implications of the measurable input variation on the development of autologous and allogeneic cell-based therapy manufacturing processes. The range of cumulative population doublings across the five BM-hMSC lines over 30 days of culture was 5.93, with an 18.2% range in colony forming efficiency at the end of the culture process and a 55.1% difference in the production of interleukin-6 between these cell lines. It has been demonstrated that this variation results in a range in the process time between these donor hMSC lines for a hypothetical product of over 13 days, creating potential batch timing issues when manufacturing products from multiple patients. All BM-hMSC donor lines demonstrated conformity to the ISCT criteria but showed a difference in cell morphology. Metabolite analysis showed that hMSCs from the different donors have a range in glucose consumption of 26.98 pmol cell(-1) day(-1), Lactate production of 29.45 pmol cell(-1) day(-1) and ammonium production of 1.35 pmol cell(-1) day(-1), demonstrating the extent of donor variability throughout the expansion process. Measuring informative product attributes during process development will facilitate progress towards consistent manufacturing processes, a critical step in the translation cell-based therapies. (C) 2015 The Authors. Published by Elsevier B.V.