Document Detail


Inoculation and growth conditions for high-cell-density expansion of mammalian neural stem cells in suspension bioreactors.
MedLine Citation:
PMID:  10099628     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Inoculation and growth conditions for the large-scale expansion of mammalian neural stem cells (NSC) have been determined. We examined suspension culture bioreactors of murine NSC, and concluded that the oxygen level should be kept high (20%), and the osmolarity of the medium should be kept low (below 400 mOsm/kg). The pH of the medium was found to have a large effect on cell proliferation, and the best growth characteristics were obtained within an optimum pH range of 7. 1 to 7.5. The inoculation conditions were also seen to have a large effect not only on the growth characteristics, but also on the number of cells that die in the initial stages of the culture. For large expansion of cells, low inoculum levels (10(4) cells/mL) and single-cell suspensions proved superior, whereas, for fast expansion of cells, higher inoculum levels (10(5) cells/mL) and spheroid inoculum forms were preferred. The inoculum temperature of the medium did not have a large effect on growth characteristics, but the pH greatly influenced cell proliferation. Inoculum pH levels should also be kept between 7.1 and 7.5. If these protocols are followed, high multiplication ratios and viabilities can be obtained in a 5-day batch suspension culture bioreactor run. A large number of cells could then be used in animal models for testing of neural drugs and in research and development toward cures for neurodegenerative disorders such as multiple sclerosis (MS) and Huntington's and Parkinson's disease. The results presented here also point the way toward studies on in vitro expansion of human neural stem cells.
Authors:
M S Kallos; L A Behie
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  63     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  1999 May 
Date Detail:
Created Date:  1999-04-26     Completed Date:  1999-04-26     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  473-83     Citation Subset:  IM    
Copyright Information:
Copyright 1999 John Wiley & Sons, Inc.
Affiliation:
Pharmaceutical Production Research Facility (PPRF), Faculty of Engineering, University of Calgary, Calgary, Alberta T2N 1N4, Canada.
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MeSH Terms
Descriptor/Qualifier:
Animals
Bioreactors
Biotechnology / instrumentation,  methods
Cell Culture Techniques / instrumentation,  methods
Cell Differentiation
Cell Division
Cell Line
Cell Transplantation
Humans
Mammals
Mice
Neurodegenerative Diseases / therapy
Neurons / cytology*
Stem Cells / cytology*

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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