Adaptation of mammalian cells to growth in serum-free media
A three-step protocol is described for adapting an anchorage-dependent, serum-dependent recombinant mammalian cell lineage to high density serum-free suspension culture. The objective is a cell lineage that is well-suited for the manufacture of a recombinant protein. The first step of the protocol g...
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| Veröffentlicht in: | Molecular biotechnology 2000-07, Vol.15 (3), p.249-257 |
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| creator | SINACORE, M. S DRAPEAU, D ADAMSON, S. R |
| description | A three-step protocol is described for adapting an anchorage-dependent, serum-dependent recombinant mammalian cell lineage to high density serum-free suspension culture. The objective is a cell lineage that is well-suited for the manufacture of a recombinant protein. The first step of the protocol generates an anchorage-independent cell lineage by culturing trypsin-treated cells in spinner flasks using serum-containing medium. The second step adapts the lineage to serum-free medium through a series of serum reduction steps in the presence of defined growth-promoting additives. The third step adapts the lineage to high-cell-density conditions by culturing the cells in a bioreactor in a manner that allows development of tolerance to growth-inhibiting substances released by the cells. Examples are presented for the use of this protocol for recombinant CHO cells. |
| doi_str_mv | 10.1385/MB:15:3:249 |
| format | Article |
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S ; DRAPEAU, D ; ADAMSON, S. R</creator><creatorcontrib>SINACORE, M. S ; DRAPEAU, D ; ADAMSON, S. R</creatorcontrib><description>A three-step protocol is described for adapting an anchorage-dependent, serum-dependent recombinant mammalian cell lineage to high density serum-free suspension culture. The objective is a cell lineage that is well-suited for the manufacture of a recombinant protein. The first step of the protocol generates an anchorage-independent cell lineage by culturing trypsin-treated cells in spinner flasks using serum-containing medium. The second step adapts the lineage to serum-free medium through a series of serum reduction steps in the presence of defined growth-promoting additives. The third step adapts the lineage to high-cell-density conditions by culturing the cells in a bioreactor in a manner that allows development of tolerance to growth-inhibiting substances released by the cells. Examples are presented for the use of this protocol for recombinant CHO cells.</description><identifier>ISSN: 1073-6085</identifier><identifier>EISSN: 1559-0305</identifier><identifier>EISSN: 1073-6085</identifier><identifier>DOI: 10.1385/MB:15:3:249</identifier><identifier>PMID: 10986701</identifier><identifier>CODEN: MLBOEO</identifier><language>eng</language><publisher>Totowa, NJ: Humana Press</publisher><subject>Adaptation ; Adaptation, Physiological ; Animal cells ; Animals ; Biological and medical sciences ; Bioreactors ; Biotechnology ; Cell Count ; Cell Culture Techniques - instrumentation ; Cell Culture Techniques - methods ; Cell Division ; Cells ; Cells, Cultured - cytology ; Cells, Cultured - physiology ; CHO Cells ; Cricetinae ; Culture Media, Serum-Free ; Establishment of new cell lines, improvement of cultural methods, mass cultures ; Eukaryotic cell cultures ; Fundamental and applied biological sciences. Psychology ; Mammals ; Methods. Procedures. 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S</creatorcontrib><creatorcontrib>DRAPEAU, D</creatorcontrib><creatorcontrib>ADAMSON, S. R</creatorcontrib><title>Adaptation of mammalian cells to growth in serum-free media</title><title>Molecular biotechnology</title><addtitle>Mol Biotechnol</addtitle><description>A three-step protocol is described for adapting an anchorage-dependent, serum-dependent recombinant mammalian cell lineage to high density serum-free suspension culture. The objective is a cell lineage that is well-suited for the manufacture of a recombinant protein. The first step of the protocol generates an anchorage-independent cell lineage by culturing trypsin-treated cells in spinner flasks using serum-containing medium. The second step adapts the lineage to serum-free medium through a series of serum reduction steps in the presence of defined growth-promoting additives. The third step adapts the lineage to high-cell-density conditions by culturing the cells in a bioreactor in a manner that allows development of tolerance to growth-inhibiting substances released by the cells. Examples are presented for the use of this protocol for recombinant CHO cells.</description><subject>Adaptation</subject><subject>Adaptation, Physiological</subject><subject>Animal cells</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Bioreactors</subject><subject>Biotechnology</subject><subject>Cell Count</subject><subject>Cell Culture Techniques - instrumentation</subject><subject>Cell Culture Techniques - methods</subject><subject>Cell Division</subject><subject>Cells</subject><subject>Cells, Cultured - cytology</subject><subject>Cells, Cultured - physiology</subject><subject>CHO Cells</subject><subject>Cricetinae</subject><subject>Culture Media, Serum-Free</subject><subject>Establishment of new cell lines, improvement of cultural methods, mass cultures</subject><subject>Eukaryotic cell cultures</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Mammals</subject><subject>Methods. Procedures. 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S</au><au>DRAPEAU, D</au><au>ADAMSON, S. R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Adaptation of mammalian cells to growth in serum-free media</atitle><jtitle>Molecular biotechnology</jtitle><addtitle>Mol Biotechnol</addtitle><date>2000-07-01</date><risdate>2000</risdate><volume>15</volume><issue>3</issue><spage>249</spage><epage>257</epage><pages>249-257</pages><issn>1073-6085</issn><eissn>1559-0305</eissn><eissn>1073-6085</eissn><coden>MLBOEO</coden><abstract>A three-step protocol is described for adapting an anchorage-dependent, serum-dependent recombinant mammalian cell lineage to high density serum-free suspension culture. The objective is a cell lineage that is well-suited for the manufacture of a recombinant protein. The first step of the protocol generates an anchorage-independent cell lineage by culturing trypsin-treated cells in spinner flasks using serum-containing medium. 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| subjects | Adaptation Adaptation, Physiological Animal cells Animals Biological and medical sciences Bioreactors Biotechnology Cell Count Cell Culture Techniques - instrumentation Cell Culture Techniques - methods Cell Division Cells Cells, Cultured - cytology Cells, Cultured - physiology CHO Cells Cricetinae Culture Media, Serum-Free Establishment of new cell lines, improvement of cultural methods, mass cultures Eukaryotic cell cultures Fundamental and applied biological sciences. Psychology Mammals Methods. Procedures. Technologies Molecular biology Proteins Recombinant Proteins - biosynthesis serum-free media Studies |
| title | Adaptation of mammalian cells to growth in serum-free media |
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