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Epithelial-mesenchymal transition in rhesus monkey embryonic stem cell colonies: the role of culturing conditions
Colonies of rhesus monkey embryonic stem cells (rhESC; cell line R366.4) have been described before to show a spatially ordered process of epithelial-mesenchymal transition in vitro. In the present investigations, we have studied variables of culturing conditions which influence the reproducibility...
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Published in: | In vitro cellular & developmental biology. Animal 2010-06, Vol.46 (6), p.516-528 |
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container_title | In vitro cellular & developmental biology. Animal |
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creator | Maranca-Hüwel, Birgit Denker, Hans-Werner |
description | Colonies of rhesus monkey embryonic stem cells (rhESC; cell line R366.4) have been described before to show a spatially ordered process of epithelial-mesenchymal transition in vitro. In the present investigations, we have studied variables of culturing conditions which influence the reproducibility of the formation of crater-like ingression centers in the colonies. Critical parameters are found to be age and density of mouse embryonic fibroblast (MEF) feeder cell layers, the mode of mitotic inactivation of the MEFs (mitomycin C, or irradiation), and the mode of rhESC isolation during subculturing (enzymatic/mechanical cell cluster isolation; type of enzyme). The described culturing system appears to offer a reproducible in vitro model potentially useful for studies on cellular processes involved in gastrulation in the primate. |
doi_str_mv | 10.1007/s11626-010-9272-8 |
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In the present investigations, we have studied variables of culturing conditions which influence the reproducibility of the formation of crater-like ingression centers in the colonies. Critical parameters are found to be age and density of mouse embryonic fibroblast (MEF) feeder cell layers, the mode of mitotic inactivation of the MEFs (mitomycin C, or irradiation), and the mode of rhESC isolation during subculturing (enzymatic/mechanical cell cluster isolation; type of enzyme). The described culturing system appears to offer a reproducible in vitro model potentially useful for studies on cellular processes involved in gastrulation in the primate.</description><identifier>ISSN: 1071-2690</identifier><identifier>EISSN: 1543-706X</identifier><identifier>DOI: 10.1007/s11626-010-9272-8</identifier><identifier>PMID: 20135359</identifier><identifier>CODEN: IVCAED</identifier><language>eng</language><publisher>New York: New York : Springer-Verlag</publisher><subject>Actins ; Age ; Animal Genetics and Genomics ; Animals ; Biomedical and Life Sciences ; CELL AND TISSUE MODELS ; Cell Biology ; Cell Culture ; Cell Culture Techniques ; Cell Differentiation ; Cell lines ; Cells, Cultured ; Colonies ; Cultured cells ; Developmental Biology ; Embryo cells ; Embryo fibroblasts ; Embryo, Mammalian - metabolism ; Embryonic stem cells ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - metabolism ; Embryos ; Enzymes ; Epithelial Cells - cytology ; Epithelial Cells - metabolism ; Feeder cells ; Fibroblasts - cytology ; Fibroblasts - metabolism ; Gastrulation ; Irradiation ; Life Sciences ; Macaca mulatta ; Mesenchymal Stromal Cells - cytology ; Mesenchymal Stromal Cells - metabolism ; Mice ; Mitomycin C ; Primates ; Radiation ; Stem Cells</subject><ispartof>In vitro cellular & developmental biology. 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Animal</title><addtitle>In Vitro Cell.Dev.Biol.-Animal</addtitle><addtitle>In Vitro Cell Dev Biol Anim</addtitle><description>Colonies of rhesus monkey embryonic stem cells (rhESC; cell line R366.4) have been described before to show a spatially ordered process of epithelial-mesenchymal transition in vitro. In the present investigations, we have studied variables of culturing conditions which influence the reproducibility of the formation of crater-like ingression centers in the colonies. Critical parameters are found to be age and density of mouse embryonic fibroblast (MEF) feeder cell layers, the mode of mitotic inactivation of the MEFs (mitomycin C, or irradiation), and the mode of rhESC isolation during subculturing (enzymatic/mechanical cell cluster isolation; type of enzyme). The described culturing system appears to offer a reproducible in vitro model potentially useful for studies on cellular processes involved in gastrulation in the primate.</description><subject>Actins</subject><subject>Age</subject><subject>Animal Genetics and Genomics</subject><subject>Animals</subject><subject>Biomedical and Life Sciences</subject><subject>CELL AND TISSUE MODELS</subject><subject>Cell Biology</subject><subject>Cell Culture</subject><subject>Cell Culture Techniques</subject><subject>Cell Differentiation</subject><subject>Cell lines</subject><subject>Cells, Cultured</subject><subject>Colonies</subject><subject>Cultured cells</subject><subject>Developmental Biology</subject><subject>Embryo cells</subject><subject>Embryo fibroblasts</subject><subject>Embryo, Mammalian - metabolism</subject><subject>Embryonic stem cells</subject><subject>Embryonic Stem Cells - cytology</subject><subject>Embryonic Stem Cells - metabolism</subject><subject>Embryos</subject><subject>Enzymes</subject><subject>Epithelial Cells - cytology</subject><subject>Epithelial Cells - metabolism</subject><subject>Feeder cells</subject><subject>Fibroblasts - cytology</subject><subject>Fibroblasts - metabolism</subject><subject>Gastrulation</subject><subject>Irradiation</subject><subject>Life Sciences</subject><subject>Macaca mulatta</subject><subject>Mesenchymal Stromal Cells - cytology</subject><subject>Mesenchymal Stromal Cells - metabolism</subject><subject>Mice</subject><subject>Mitomycin C</subject><subject>Primates</subject><subject>Radiation</subject><subject>Stem Cells</subject><issn>1071-2690</issn><issn>1543-706X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><recordid>eNqFkctu1TAQhiMEoqXwACwAiw2rgG_xpTtUlYtUiQVUYmc5jtPjg2OfepLFeXscUorEAryxNfPNP575m-Y5wW8JxvIdECKoaDHBraaStupBc0o6zlqJxfeH9Y0laanQ-KR5ArDH9WgiHjcnFBPWsU6fNreXhzDvfAw2tpMHn9zuONmI5mIThDnkhEJCZedhATTl9MMfkZ_6cswpOASzn5DzMSKXY414OEdVDZUcPcojckuclxLSTc2n4ZccPG0ejTaCf3Z3nzXXHy6_XXxqr758_Hzx_qp1nKu5ZZ0aiKKYjV4OWFlrBRHW9VJT1nXcupHLvldSS2eF9MwNwnHcD3UxgxDWsrPmzaZ7KPl28TCbKcD6V5t8XsBoTJmUSrH_kpKxujfOu0q-_ovc56WkOoYRWGvZCS4qRDbIlQxQ_GgOJUy2HA3BZvXNbL6Z6ptZfTOq1ry8E176yQ_3Fb-NqgDdADis-_TlT-d_qb7YivYw53IvyrGqs2tS86-2_GizsTclgLn-urbERHFVG7OfRa-3KQ</recordid><startdate>20100601</startdate><enddate>20100601</enddate><creator>Maranca-Hüwel, Birgit</creator><creator>Denker, Hans-Werner</creator><general>New York : Springer-Verlag</general><general>Springer Science + Business Media</general><general>Springer-Verlag</general><general>Society for In Vitro Biology</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>4T-</scope><scope>7QL</scope><scope>7T7</scope><scope>7TK</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AF</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>S0X</scope><scope>7X8</scope><scope>7QO</scope><scope>RC3</scope></search><sort><creationdate>20100601</creationdate><title>Epithelial-mesenchymal transition in rhesus monkey embryonic stem cell colonies: the role of culturing conditions</title><author>Maranca-Hüwel, Birgit ; 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Animal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Maranca-Hüwel, Birgit</au><au>Denker, Hans-Werner</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Epithelial-mesenchymal transition in rhesus monkey embryonic stem cell colonies: the role of culturing conditions</atitle><jtitle>In vitro cellular & developmental biology. Animal</jtitle><stitle>In Vitro Cell.Dev.Biol.-Animal</stitle><addtitle>In Vitro Cell Dev Biol Anim</addtitle><date>2010-06-01</date><risdate>2010</risdate><volume>46</volume><issue>6</issue><spage>516</spage><epage>528</epage><pages>516-528</pages><issn>1071-2690</issn><eissn>1543-706X</eissn><coden>IVCAED</coden><abstract>Colonies of rhesus monkey embryonic stem cells (rhESC; cell line R366.4) have been described before to show a spatially ordered process of epithelial-mesenchymal transition in vitro. In the present investigations, we have studied variables of culturing conditions which influence the reproducibility of the formation of crater-like ingression centers in the colonies. Critical parameters are found to be age and density of mouse embryonic fibroblast (MEF) feeder cell layers, the mode of mitotic inactivation of the MEFs (mitomycin C, or irradiation), and the mode of rhESC isolation during subculturing (enzymatic/mechanical cell cluster isolation; type of enzyme). The described culturing system appears to offer a reproducible in vitro model potentially useful for studies on cellular processes involved in gastrulation in the primate.</abstract><cop>New York</cop><pub>New York : Springer-Verlag</pub><pmid>20135359</pmid><doi>10.1007/s11626-010-9272-8</doi><tpages>13</tpages></addata></record> |
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subjects | Actins Age Animal Genetics and Genomics Animals Biomedical and Life Sciences CELL AND TISSUE MODELS Cell Biology Cell Culture Cell Culture Techniques Cell Differentiation Cell lines Cells, Cultured Colonies Cultured cells Developmental Biology Embryo cells Embryo fibroblasts Embryo, Mammalian - metabolism Embryonic stem cells Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Embryos Enzymes Epithelial Cells - cytology Epithelial Cells - metabolism Feeder cells Fibroblasts - cytology Fibroblasts - metabolism Gastrulation Irradiation Life Sciences Macaca mulatta Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - metabolism Mice Mitomycin C Primates Radiation Stem Cells |
title | Epithelial-mesenchymal transition in rhesus monkey embryonic stem cell colonies: the role of culturing conditions |
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