Document Detail


Good manufacturing practice-compliant expansion of marrow-derived stem and progenitor cells for cell therapy.
MedLine Citation:
PMID:  18019358     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Ex vivo expansion is being used to increase the number of stem and progenitor cells for autologous cell therapy. Initiation of pivotal clinical trials testing the efficacy of these cells for tissue repair has been hampered by the challenge of assuring safe and high-quality cell production. A strategy is described here for clinical-scale expansion of bone marrow (BM)-derived stem cells within a mixed cell population in a completely closed process from cell collection through postculture processing using sterile connectable devices. Human BM mononuclear cells (BMMNC) were isolated, cultured for 12 days, and washed postharvest using either standard open procedures in laminar flow hoods or using automated closed systems. Conditions for these studies were similar to long-term BM cultures in which hematopoietic and stromal components are cultured together. Expansion of marrow-derived stem and progenitor cells was then assessed. Cell yield, number of colony forming units (CFU), phenotype, stability, and multilineage differentiation capacity were compared from the single pass perfusion bioreactor and standard flask cultures. Purification of BMMNC using a closed Ficoll gradient process led to depletion of 98% erythrocytes and 87% granulocytes, compared to 100% and 70%, respectively, for manual processing. After closed system culture, mesenchymal progenitors, measured as CD105+CD166+CD14-CD45- and fibroblastic CFU, expanded 317- and 364-fold, respectively, while CD34+ hematopoietic progenitors were depleted 10-fold compared to starting BMMNC. Cultured cells exhibited multilineage differentiation by displaying adipogenic, osteogenic, and endothelial characteristics in vitro. No significant difference was observed between manual and bioreactor cultures. Automated culture and washing of the cell product resulted in 181 x 10(6) total cells that were viable and contained fibroblastic CFU for at least 24 h of storage. A combination of closed, automated technologies enabled production of good manufacturing practice (GMP)-compliant cell therapeutics, ready for use within a clinical setting, with minimal risk of microbial contamination.
Authors:
Martin H Gastens; Kristin Goltry; Wolfgang Prohaska; Diethelm Tschöpe; Bernd Stratmann; Dirk Lammers; Stanley Kirana; Christian Götting; Knut Kleesiek
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cell transplantation     Volume:  16     ISSN:  0963-6897     ISO Abbreviation:  Cell Transplant     Publication Date:  2007  
Date Detail:
Created Date:  2007-11-16     Completed Date:  2008-02-04     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9208854     Medline TA:  Cell Transplant     Country:  United States    
Other Details:
Languages:  eng     Pagination:  685-96     Citation Subset:  IM    
Affiliation:
Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany. gastens@gmx.de
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MeSH Terms
Descriptor/Qualifier:
Bone Marrow Cells / cytology,  physiology*
Cell Culture Techniques* / instrumentation
Cell Lineage
Colony-Forming Units Assay
Guideline Adherence
Humans
Stem Cell Transplantation*
Stem Cells / cytology,  physiology*
Tissue Therapy* / instrumentation,  methods,  standards

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