This title appears in the Scientific Report :
2005
Please use the identifier:
http://hdl.handle.net/2128/2471 in citations.
Ex-vivo Generierung von neutrophilen Zellen zur Prävention und Therapie der Sepsis
Ex-vivo Generierung von neutrophilen Zellen zur Prävention und Therapie der Sepsis
Neutrophil granulocytes represent the key element of native immunity against microbial infection. Failure of this system leads to a whole body infection and as a consequence thereof to life-threatening symptoms of sepsis. In this work strategies and systems were developed for cell based prevention a...
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Personal Name(s): | Herbold, Ralf (Corresponding author) |
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Contributing Institute: |
Biotechnologie 2; IBT-2 |
Imprint: |
Jülich
Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag
2005
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Dissertation Note: |
Universität Bonn, Diss., 2005 |
ISBN: |
3-89336-407-2 |
Document Type: |
Book Dissertation / PhD Thesis |
Research Program: |
Biotechnologie |
Series Title: |
Schriften des Forschungszentrums Jülich. Reihe Lebenswissenschaften / Life Sciences
18 |
Subject (ZB): | |
Link: |
OpenAccess |
Publikationsportal JuSER |
Neutrophil granulocytes represent the key element of native immunity against microbial infection. Failure of this system leads to a whole body infection and as a consequence thereof to life-threatening symptoms of sepsis. In this work strategies and systems were developed for cell based prevention and therapy of sepsis. The developed Mini-Spinner system allows for the first time the multi-parallel cultivation of both cell lines and primary cells under fully controlled conditions in volumes of 20 to 50 mL. Using this system a clinically applicable cultivation process for the ex-vivo generation of neutrophil granulocytes starting from hematopoeietic stem and progenitor cells was developed. Medium composition, growth factor combination as well as relevant ambient conditions like temperature, pH and pO$_{2}$ were optimized. The proportion of neutrophil cell types exceeded 75% of total cell count by day 10. At day 16 cells showed the typical banded and segmented morphology as well as phagocytosis and oxidative-burst activity which are typical for mature neutrohil granulocytes. In this thesis a scalable process for the production and maturation of HL-60 cells was developed allowing cost-effective production of cells with neutrophil functionality in large quantities. This might be useful for the therapy of acute septic events as the generated cells can be stored until needed. In a first step relevant process parameters (medium, supplements, ambient parameters, feeding) were optimized for the expansion of HL-60 biomass. Maturation of HL-60 is induced using ATRA at concentrations from 30 to 50 μM during a time-span of 22.5 to 48 h. Both processes allow the ex-vivo generation of neutrophil granulocytes for cell-based prevention ant therapy of sepsis and might be reasonable alternatives for the usage of donor-granulocytes. |