Expansion, harvest and cryopreservation of human mesenchymal stem cells in a serum-free microcarrier process.
Thomas R.J. Heathman,Veronica A.M. Glyn,Andrew Picken,Qasim A. Rafiq,Qasim A. Rafiq,Karen Coopman,Alvin W. Nienow,Alvin W. Nienow,Bo Kara,Christopher J. Hewitt,Christopher J. Hewitt +10 more
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TLDR
This approach demonstrates that once an h MSC 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.Abstract:
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 100mL stirred spinner flasks at a density of 3×105cells.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 3h 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.read more
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Manufacturing human mesenchymal stem cells at clinical scale: Process and regulatory challenges
TL;DR: This mini-review summarizes both the current state of the hMSC production process and the challenges which have to be taken into account when efficiently producing hMSCs for the clinical scale and special emphasis is placed on the upstream processing (USP) and DSP operations which cover expansion, harvesting, detachment, separation, washing and concentration steps.
Journal ArticleDOI
Bioprocessing strategies for the large-scale production of human mesenchymal stem cells: a review.
TL;DR: Optimize a bioprocess to generate hMSCs or their secreted products (or both) promises to improve the efficacy as well as safety of this stem cell therapy.
Journal ArticleDOI
Characterization of human mesenchymal stem cells from multiple donors and the implications for large scale bioprocess development
Thomas R.J. Heathman,Qasim A. Rafiq,Qasim A. Rafiq,Alexander K. C. Chan,Karen Coopman,Alvin W. Nienow,Alvin W. Nienow,Bo Kara,Christopher J. Hewitt,Christopher J. Hewitt +9 more
TL;DR: In this article, the authors 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.
Journal ArticleDOI
Metabolism in Human Mesenchymal Stromal Cells: A Missing Link Between hMSC Biomanufacturing and Therapy?
TL;DR: Understanding the regulatory mechanisms underpinning hMSC phenotypic and functional property is crucial for developing novel engineering protocols that maximize yield while preserving therapeutic potency in hMSCs clinical translation.
Journal ArticleDOI
Agitation conditions for the culture and detachment of hMSCs from microcarriers in multiple bioreactor platforms
Alvin W. Nienow,Alvin W. Nienow,Christopher J. Hewitt,Christopher J. Hewitt,Thomas R.J. Heathman,Veronica A.M. Glyn,Gonҫalo N. Fonte,Mariana P. Hanga,Karen Coopman,Qasim A. Rafiq,Qasim A. Rafiq +10 more
TL;DR: A scaleable protocol for the detachment from microcarriers in spinner flasks of hMSCs from two donors and cells were shown to retain their desired quality attributes and were able to proliferate, offers a sound basis for a wide range of scales of operation.
References
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Journal ArticleDOI
Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement
Massimo Dominici,K. Le Blanc,Ingo Mueller,I. Slaper-Cortenbach,Frank C. Marini,Diane S. Krause,Robert J. Deans,Armand Keating,Darwin J. Prockop,Edwin M. Horwitz +9 more
TL;DR: The Mesenchymal and Tissue Stem Cell Committee of the International Society for Cellular Therapy proposes minimal criteria to define human MSC, believing this minimal set of standard criteria will foster a more uniform characterization of MSC and facilitate the exchange of data among investigators.
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Mesenchymal stem cells
TL;DR: The study of mesenchymal stem cells, whether isolated from embryos or adults, provides the basis for the emergence of a new therapeutic technology of self‐cell repair.
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Mesenchymal Stem Cells
TL;DR: The bone marrow contains multipotent MSC, which can be easily isolated and cultured in vitro, and the possibility of their clinical use in cell and gene therapy is analyzed.
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Mesenchymal stem cells as trophic mediators.
Arnold I. Caplan,James E. Dennis +1 more
TL;DR: Several studies which tested the use of MSCs in models of infarct (injured heart), stroke (brain), or meniscus regeneration models are reviewed within the context of M SC‐mediated trophic effects in tissue repair.
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A Randomized, Double-Blind, Placebo-Controlled, Dose-Escalation Study of Intravenous Adult Human Mesenchymal Stem Cells (Prochymal) After Acute Myocardial Infarction
Joshua M. Hare,Jay H. Traverse,Timothy D. Henry,Nabil Dib,Robert K. Strumpf,Steven P. Schulman,Gary Gerstenblith,Anthony N. DeMaria,Ali E. Denktas,Roger Gammon,James B. Hermiller,Mark A. Reisman,Gary L. Schaer,Warren Sherman +13 more
TL;DR: This trial provides pivotal safety and provisional efficacy data for an allogeneic bone marrow-derived stem cell in post-infarction patients and indicates intravenousAllogeneic hMSCs are safe in patients after acute MI.