Inoculation and growth conditions for high‐cell‐density expansion of mammalian neural stem cells in suspension bioreactors
Identifieur interne : 001802 ( Istex/Corpus ); précédent : 001801; suivant : 001803Inoculation and growth conditions for high‐cell‐density expansion of mammalian neural stem cells in suspension bioreactors
Auteurs : Michael S. Kallos ; Leo A. BehieSource :
- Biotechnology and Bioengineering [ 0006-3592 ] ; 1999-05-20.
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- KwdEn :
Abstract
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 (104 cells/mL) and single‐cell suspensions proved superior, whereas, for fast expansion of cells, higher inoculum levels (105 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. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 473–483, 1999.
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DOI: 10.1002/(SICI)1097-0290(19990520)63:4<473::AID-BIT11>3.0.CO;2-C
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ISTEX:29F3878322289E4EA4B0021B07E1CDB3BC3D94BBLe document en format XML
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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 (104 cells/mL) and single‐cell suspensions proved superior, whereas, for fast expansion of cells, higher inoculum levels (105 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. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 473–483, 1999.</div></front></TEI><istex><corpusName>wiley</corpusName><author><json:item><name>Michael S. Kallos</name><affiliations><json:string>Pharmaceutical Production Research Facility (PPRF), Faculty of Engineering, University of Calgary, Calgary, Alberta T2N 1N4, Canada</json:string></affiliations></json:item><json:item><name>Leo A. 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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<sup>4</sup> cells/mL) and single‐cell suspensions proved superior, whereas, for fast expansion of cells, higher inoculum levels (10<sup>5</sup> 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. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 473–483, 1999.</p></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Inoculation and growth conditions for high‐cell‐density expansion of mammalian neural stem cells in suspension bioreactors</title></titleInfo><titleInfo type="abbreviated" lang="en"><title>MAMMALIAN NEURAL STEM CELLS IN SUSPENSION BIOREACTORS</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Inoculation and growth conditions for high‐cell‐density expansion of mammalian neural stem cells in suspension bioreactors</title></titleInfo><name type="personal"><namePart type="given">Michael S.</namePart><namePart type="family">Kallos</namePart><affiliation>Pharmaceutical Production Research Facility (PPRF), Faculty of Engineering, University of Calgary, Calgary, Alberta T2N 1N4, Canada</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Leo A.</namePart><namePart type="family">Behie</namePart><affiliation>Pharmaceutical Production Research Facility (PPRF), Faculty of Engineering, University of Calgary, Calgary, Alberta T2N 1N4, Canada</affiliation><affiliation>Pharmaceutical Production Research Facility (PPRF), Faculty of Engineering, University of Calgary, Calgary, Alberta T2N 1N4, Canada</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="article"></genre><originInfo><publisher>John Wiley & Sons, Inc.</publisher><place><placeTerm type="text">New York</placeTerm></place><dateIssued encoding="w3cdtf">1999-05-20</dateIssued><dateCaptured encoding="w3cdtf">1998-07-24</dateCaptured><dateValid encoding="w3cdtf">1998-11-06</dateValid><copyrightDate encoding="w3cdtf">1999</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType><extent unit="figures">12</extent><extent unit="tables">1</extent><extent unit="references">33</extent><extent unit="words">7352</extent></physicalDescription><abstract lang="en">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 (104 cells/mL) and single‐cell suspensions proved superior, whereas, for fast expansion of cells, higher inoculum levels (105 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. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 473–483, 1999.</abstract><note type="funding">NSERC of Canada</note><subject lang="en"><genre>keywords</genre><topic>mammalian neural stem cells</topic><topic>aggregates</topic><topic>suspension bioreactor</topic><topic>growth conditions</topic><topic>inoculation conditions</topic><topic>expansion</topic></subject><relatedItem type="host"><titleInfo><title>Biotechnology and Bioengineering</title></titleInfo><titleInfo type="abbreviated"><title>Biotechnol. Bioeng.</title></titleInfo><genre type="journal">journal</genre><identifier type="ISSN">0006-3592</identifier><identifier type="eISSN">1097-0290</identifier><identifier type="DOI">10.1002/(ISSN)1097-0290</identifier><identifier type="PublisherID">BIT</identifier><part><date>1999</date><detail type="volume"><caption>vol.</caption><number>63</number></detail><detail type="issue"><caption>no.</caption><number>4</number></detail><extent unit="pages"><start>473</start><end>483</end><total>11</total></extent></part></relatedItem><identifier type="istex">29F3878322289E4EA4B0021B07E1CDB3BC3D94BB</identifier><identifier type="DOI">10.1002/(SICI)1097-0290(19990520)63:4<473::AID-BIT11>3.0.CO;2-C</identifier><identifier type="ArticleID">BIT11</identifier><accessCondition type="use and reproduction" contentType="copyright">Copyright © 1999 John Wiley & Sons, Inc.</accessCondition><recordInfo><recordContentSource>WILEY</recordContentSource><recordOrigin>John Wiley & Sons, Inc.</recordOrigin></recordInfo></mods></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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