Expansion, harvest and cryopreservation of human mesenchymal stem cells in a serum-free microcarrier process.
Expansion, harvest and cryopreservation of human mesenchymal stem cells in a serum-free microcarrier process.
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DOI：
10.1002/bit.25582
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发表时间：
2015-08
影响因子：
3.8
通讯作者：
Hewitt, Christopher J.
中科院分区：
文献类型：
--
作者：
Heathman, Thomas R. J.;Glyn, Veronica A. M.;Picken, Andrew;Rafiq, Qasim A.;Coopman, Karen;Nienow, Alvin W.;Kara, Bo;Hewitt, Christopher J.
关键词：
人类间充质干细胞 (hMSC) 疗法目前正在临床开发中取得进展，推动了对一致且具有成本效益的制造工艺的需求，以满足商业生产所需的批量大小。动物源性血清在 hMSC 培养中很常见，但存在许多缺点，例如供应有限、批次间差异、监管负担增加、病原体传播的可能性以及工艺优化范围缩小。这些限制可能会影响一致的大规模工艺的开发，因此必须得到解决。因此，这项工作的目的是对无血清 hMSC 制造工艺的系统开发进行试点研究。人骨髓来源的 hMSC 在 100 mL 搅拌旋转烧瓶中的纤连蛋白包被的无孔塑料微载体上以 3××105 个细胞.mL−1 的密度在无血清培养基中扩增。通过我们最近开发的技术，使用非动物酶促细胞分离并搅拌，然后过滤以将 hMSC 与微载体分离，成功收获了 hMSC，收获后活力为 99.63±0.03%。发现 hMSC 符合 ISCT 表征标准，并保持 hMSC 生长和集落形成潜力。 hMSC 收获后悬浮，以模拟规模化扩增过程的典型汇集时间，并使用可控速率冷冻过程冷冻保存在无血清载体溶液中。解冻后活力为 75.8±1.4%，也观察到类似的 3 小时附着效率，表明 hMSC 恢复和附着成功。因此，这种方法表明，一旦选择了 hMSC 系和适当的培养基进行生产，就可以整合多个单元操作，以生成从扩增到冷冻保存的无动物成分的 hMSC 生产过程。生物技术。生物工程。 2015；112：1696–1707。 © 2015 作者。生物技术和生物工程由 Wiley periodicals, Inc. 出版
Human mesenchymal stem cell (hMSC) therapies are currently progressing through clinical development, driving the need for consistent, and cost effective manufacturing processes to meet the lot‐sizes required for commercial production. The use of animal‐derived serum is common in hMSC culture but has many drawbacks such as limited supply, lot‐to‐lot variability, increased regulatory burden, possibility of pathogen transmission, and reduced scope for process optimization. These constraints may impact the development of a consistent large‐scale process and therefore must be addressed. The aim of this work was therefore to run a pilot study in the systematic development of serum‐free hMSC manufacturing process. Human bone‐marrow derived hMSCs were expanded on fibronectin‐coated, non‐porous plastic microcarriers in 100 mL stirred spinner flasks at a density of 3 × 105 cells.mL−1 in serum‐free medium. The hMSCs were successfully harvested by our recently‐developed technique using animal‐free enzymatic cell detachment accompanied by agitation followed by filtration to separate the hMSCs from microcarriers, with a post‐harvest viability of 99.63 ± 0.03%. The hMSCs were found to be in accordance with the ISCT characterization criteria and maintained hMSC outgrowth and colony‐forming potential. The hMSCs were held in suspension post‐harvest to simulate a typical pooling time for a scaled expansion process and cryopreserved in a serum‐free vehicle solution using a controlled‐rate freezing process. Post‐thaw viability was 75.8 ± 1.4% with a similar 3 h attachment efficiency also observed, indicating successful hMSC recovery, and attachment. This approach therefore demonstrates that once an hMSC line and appropriate medium have been selected for production, multiple unit operations can be integrated to generate an animal component‐free hMSC production process from expansion through to cryopreservation. Biotechnol. Bioeng. 2015;112: 1696–1707. © 2015 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.
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影响因子：
24
作者：
Hare, Joshua M.;Traverse, Jay H.;Henry, Timothy D.;Dib, Nabil;Strumpf, Robert K.;Schulman, Steven P.;Gerstenblith, Gary;DeMaria, Anthony N.;Denktas, Ali E.;Gammon, Roger S.;Hermiller, James B., Jr.;Reisman, Mark A.;Schaer, Gary L.;Sherman, Warren
通讯作者：
Sherman, Warren
DOI：
10.1186/1478-811x-8-18
发表时间：
2010-07-16
期刊：
Cell communication and signaling : CCS
影响因子：
--
作者：
Lavrentieva A;Majore I;Kasper C;Hass R
通讯作者：
Hass R
影响因子：
4
作者：
Dreher, Lena;Elvers-Hornung, Susanne;Bieback, Karen
通讯作者：
Bieback, Karen
影响因子：
4
作者：
Caplan, Arnold I.;Dennis, James E.
通讯作者：
Dennis, James E.
影响因子：
2.8
作者：
CAPLAN, AI
通讯作者：
CAPLAN, AI