Autologous Feeder Cells from Embryoid Body Outgrowth Support the Long-Term Growth of Human Embryonic Stem Cells More Effectively than Those from Direct Differentiation
Autologous feeder cells have been developed by various methods to minimize the presence of xenogenic entities in human embryonic stem cell (hESC) cultures. However, there was no systematic comparison of supportive effects of the feeder cells on hESC growth, nor comparison to the supportive effects o...
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| Veröffentlicht in: | Tissue engineering. Part C, Methods Methods, 2010-08, Vol.16 (4), p.719-733 |
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| creator | Fu, Xin Toh, Wei Seong Liu, Hua Lu, Kai Li, Mingming Hande, Manoor Prakash Cao, Tong |
| description | Autologous feeder cells have been developed by various methods to minimize the presence of xenogenic entities in human embryonic stem cell (hESC) cultures. However, there was no systematic comparison of supportive effects of the feeder cells on hESC growth, nor comparison to the supportive effects of various feeder-free culture systems and standard mouse feeder cells. In this study, we aimed to compare the supportive abilities of autologous feeders derived either directly from H9 hESCs (H9 dF) or from outgrowth of embryoid body predifferentiated in suspension from H9 hESCs (H9 ebF). Mouse feeder system and matrigel-mTeSR1 feeder-free system were used as controls. H9 ebF was found to secrete more basic fibroblast growth factor in the conditioned medium than H9 dF did. The undifferentiated state of H9 hESCs was sustained more stably on H9 ebF than on H9 dF, and the differentiation potential of H9 hESCs on H9 ebF was higher than on H9 dF. We concluded that H9 ebF was an optimal autologous feeder to maintain the long-term undifferentiated state of hESCs in our current culture system. This study helps to standardize the autologous culture of hESCs. It also suggests a more definite direction for future development of xeno-free culture system for hESCs. |
| doi_str_mv | 10.1089/ten.tec.2009.0360 |
| format | Article |
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However, there was no systematic comparison of supportive effects of the feeder cells on hESC growth, nor comparison to the supportive effects of various feeder-free culture systems and standard mouse feeder cells. In this study, we aimed to compare the supportive abilities of autologous feeders derived either directly from H9 hESCs (H9 dF) or from outgrowth of embryoid body predifferentiated in suspension from H9 hESCs (H9 ebF). Mouse feeder system and matrigel-mTeSR1 feeder-free system were used as controls. H9 ebF was found to secrete more basic fibroblast growth factor in the conditioned medium than H9 dF did. The undifferentiated state of H9 hESCs was sustained more stably on H9 ebF than on H9 dF, and the differentiation potential of H9 hESCs on H9 ebF was higher than on H9 dF. We concluded that H9 ebF was an optimal autologous feeder to maintain the long-term undifferentiated state of hESCs in our current culture system. This study helps to standardize the autologous culture of hESCs. It also suggests a more definite direction for future development of xeno-free culture system for hESCs.</description><identifier>ISSN: 1937-3384</identifier><identifier>EISSN: 1937-3392</identifier><identifier>DOI: 10.1089/ten.tec.2009.0360</identifier><identifier>PMID: 19911961</identifier><language>eng</language><publisher>United States: Mary Ann Liebert, Inc</publisher><subject>Animals ; Cell Culture Techniques - methods ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Cells ; Collagen - metabolism ; Cryopreservation ; Culture Media, Conditioned - metabolism ; Drug Combinations ; Embryo, Mammalian - cytology ; Embryo, Mammalian - metabolism ; Embryonic stem cells ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - metabolism ; Embryos ; Fibroblast Growth Factor 2 - secretion ; Fibroblast growth factors ; Fibroblasts - cytology ; Flow Cytometry ; Gene Expression Regulation ; Growth ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Karyotyping ; Laminin - metabolism ; Mice ; Properties ; Proteoglycans - metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Teratoma - pathology ; Time Factors</subject><ispartof>Tissue engineering. 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Part C, Methods</title><addtitle>Tissue Eng Part C Methods</addtitle><description>Autologous feeder cells have been developed by various methods to minimize the presence of xenogenic entities in human embryonic stem cell (hESC) cultures. However, there was no systematic comparison of supportive effects of the feeder cells on hESC growth, nor comparison to the supportive effects of various feeder-free culture systems and standard mouse feeder cells. In this study, we aimed to compare the supportive abilities of autologous feeders derived either directly from H9 hESCs (H9 dF) or from outgrowth of embryoid body predifferentiated in suspension from H9 hESCs (H9 ebF). Mouse feeder system and matrigel-mTeSR1 feeder-free system were used as controls. H9 ebF was found to secrete more basic fibroblast growth factor in the conditioned medium than H9 dF did. The undifferentiated state of H9 hESCs was sustained more stably on H9 ebF than on H9 dF, and the differentiation potential of H9 hESCs on H9 ebF was higher than on H9 dF. We concluded that H9 ebF was an optimal autologous feeder to maintain the long-term undifferentiated state of hESCs in our current culture system. This study helps to standardize the autologous culture of hESCs. It also suggests a more definite direction for future development of xeno-free culture system for hESCs.</description><subject>Animals</subject><subject>Cell Culture Techniques - methods</subject><subject>Cell Differentiation</subject><subject>Cell Line</subject><subject>Cell Proliferation</subject><subject>Cells</subject><subject>Collagen - metabolism</subject><subject>Cryopreservation</subject><subject>Culture Media, Conditioned - metabolism</subject><subject>Drug Combinations</subject><subject>Embryo, Mammalian - cytology</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>Fibroblast Growth Factor 2 - secretion</subject><subject>Fibroblast growth factors</subject><subject>Fibroblasts - cytology</subject><subject>Flow Cytometry</subject><subject>Gene Expression Regulation</subject><subject>Growth</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>In Situ Hybridization, Fluorescence</subject><subject>Karyotyping</subject><subject>Laminin - metabolism</subject><subject>Mice</subject><subject>Properties</subject><subject>Proteoglycans - metabolism</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Teratoma - pathology</subject><subject>Time Factors</subject><issn>1937-3384</issn><issn>1937-3392</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkstu1DAYhSMEomXgAdggCxZdJfiWi5fDMG2RBnXRYW05zp8ZV4k9OA5onojXrKNEIBASyLZ-y_7O8UUnSV4TnBFcifcBbBZAZxRjkWFW4CfJJRGsTBkT9OnPecUvkhfD8IBxgYtSPE8uiBCEiIJcJj_WY3CdO7hxQNcADXi0ga4bUOtdj7Z97c_ONOiDa87obgwH776HI7ofTyfnAwpHQDtnD-kefI9u5k3XotuxV3ZRW6PRfYB-8f3sPKBt24IO5ht05-gR0f3RDTCf-dH4uBdLZDzYYFQwzr5MnrWqG-DVUlfJl-vtfnOb7u5uPm3Wu1TnHIcUaCUU01hRQfO64jqnHFRTqFKLui4LweuyJA0DCi3Xba00YwAKN6zBNVDBVsnV7Hvy7usIQ5C9GXS8ubIQ_0iWeU44JYT_m2QcE0p4Hsm3f5APbvQ2PkOWOC8oYbGvknczdFAdSGNbF7zSk6VcU15xHA8uIpX9hYqtgd5oZ6E1cf03AZkF2rth8NDKkze98mdJsJxCJGOI4tByCpGcQhQ1b5b7jnUPzS_FkpoIlDMwLStrOwM1-PAf1o8LqtgH</recordid><startdate>20100801</startdate><enddate>20100801</enddate><creator>Fu, Xin</creator><creator>Toh, Wei Seong</creator><creator>Liu, Hua</creator><creator>Lu, Kai</creator><creator>Li, Mingming</creator><creator>Hande, Manoor Prakash</creator><creator>Cao, Tong</creator><general>Mary Ann Liebert, Inc</general><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>7QP</scope><scope>7T5</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</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>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>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>RC3</scope><scope>7X8</scope><scope>7QO</scope></search><sort><creationdate>20100801</creationdate><title>Autologous Feeder Cells from Embryoid Body Outgrowth Support the Long-Term Growth of Human Embryonic Stem Cells More Effectively than Those from Direct Differentiation</title><author>Fu, Xin ; 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Part C, Methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fu, Xin</au><au>Toh, Wei Seong</au><au>Liu, Hua</au><au>Lu, Kai</au><au>Li, Mingming</au><au>Hande, Manoor Prakash</au><au>Cao, Tong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Autologous Feeder Cells from Embryoid Body Outgrowth Support the Long-Term Growth of Human Embryonic Stem Cells More Effectively than Those from Direct Differentiation</atitle><jtitle>Tissue engineering. Part C, Methods</jtitle><addtitle>Tissue Eng Part C Methods</addtitle><date>2010-08-01</date><risdate>2010</risdate><volume>16</volume><issue>4</issue><spage>719</spage><epage>733</epage><pages>719-733</pages><issn>1937-3384</issn><eissn>1937-3392</eissn><abstract>Autologous feeder cells have been developed by various methods to minimize the presence of xenogenic entities in human embryonic stem cell (hESC) cultures. However, there was no systematic comparison of supportive effects of the feeder cells on hESC growth, nor comparison to the supportive effects of various feeder-free culture systems and standard mouse feeder cells. In this study, we aimed to compare the supportive abilities of autologous feeders derived either directly from H9 hESCs (H9 dF) or from outgrowth of embryoid body predifferentiated in suspension from H9 hESCs (H9 ebF). Mouse feeder system and matrigel-mTeSR1 feeder-free system were used as controls. H9 ebF was found to secrete more basic fibroblast growth factor in the conditioned medium than H9 dF did. The undifferentiated state of H9 hESCs was sustained more stably on H9 ebF than on H9 dF, and the differentiation potential of H9 hESCs on H9 ebF was higher than on H9 dF. We concluded that H9 ebF was an optimal autologous feeder to maintain the long-term undifferentiated state of hESCs in our current culture system. This study helps to standardize the autologous culture of hESCs. It also suggests a more definite direction for future development of xeno-free culture system for hESCs.</abstract><cop>United States</cop><pub>Mary Ann Liebert, Inc</pub><pmid>19911961</pmid><doi>10.1089/ten.tec.2009.0360</doi><tpages>15</tpages></addata></record> |
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| ispartof | Tissue engineering. Part C, Methods, 2010-08, Vol.16 (4), p.719-733 |
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| language | eng |
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| source | Mary Ann Liebert Online Subscription; MEDLINE; Alma/SFX Local Collection |
| subjects | Animals Cell Culture Techniques - methods Cell Differentiation Cell Line Cell Proliferation Cells Collagen - metabolism Cryopreservation Culture Media, Conditioned - metabolism Drug Combinations Embryo, Mammalian - cytology Embryo, Mammalian - metabolism Embryonic stem cells Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Embryos Fibroblast Growth Factor 2 - secretion Fibroblast growth factors Fibroblasts - cytology Flow Cytometry Gene Expression Regulation Growth Humans Immunohistochemistry In Situ Hybridization, Fluorescence Karyotyping Laminin - metabolism Mice Properties Proteoglycans - metabolism Reverse Transcriptase Polymerase Chain Reaction Teratoma - pathology Time Factors |
| title | Autologous Feeder Cells from Embryoid Body Outgrowth Support the Long-Term Growth of Human Embryonic Stem Cells More Effectively than Those from Direct Differentiation |
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