SPARC is involved in the maintenance of mitotically inactivated mouse embryonic fibroblast cells
Identifieur interne : 002110 ( Main/Exploration ); précédent : 002109; suivant : 002111SPARC is involved in the maintenance of mitotically inactivated mouse embryonic fibroblast cells
Auteurs : Jun Yeon Won [Corée du Sud] ; Young Jin Lee [États-Unis] ; Seung-Joon Lee [Corée du Sud] ; Woo Jin Kim [Corée du Sud] ; Seon-Sook Han [Corée du Sud] ; Se-Ran Yang [Corée du Sud] ; Heung-Myong Woo [Corée du Sud] ; Sung-Min Park [Corée du Sud] ; Hyang-Ah Lee [Corée du Sud] ; Seok-Ho Hong [Corée du Sud]Source :
- In Vitro Cellular & Developmental Biology - Animal [ 1071-2690 ] ; 2013-06-01.
English descriptors
- KwdEn :
- GeneFishing, MEFs, SPARC, hPSC.
Abstract
Abstract: Mitotically inactivated feeder cells such as mouse embryonic fibroblast (MEFs) cells have been widely applied for physical and physiological support in the pluripotency maintenance of human pluripotent stem cells (hPSCs). However, accurate supporting mechanism or factors of feeder cells are poorly understood. Here, we isolated differentially expressed genes between wild-type MEFs and mitotically inactivated MEFs (miMEFs) by employing annealing control primer-based GeneFishing polymerase chain reaction. We identified a secreted protein acidic cysteine-rich glycoprotein (SPARC) gene that is upregulated in miMEFs. Suppression of SPARC expression in miMEFs using small interference RNA (siRNA) displayed gradual detachment of miMEFs. Furthermore, we found a significant reduction of OCT4- and SSEA3-positive hPS cell population maintained on SPARC siRNA-miMEFs compared to on miMEFs by flow cytometrical analysis. These findings suggest that SPARC plays a critical role in the maintenance of miMEFs without loss of cell number and might be a key component for supporting the culture of hPSCs.
Url:
DOI: 10.1007/s11626-013-9601-9
Affiliations:
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Jin" sort="Kim, Woo Jin" uniqKey="Kim W" first="Woo Jin" last="Kim">Woo Jin Kim</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Internal Medicine, School of Medicine, Kangwon National University, Hyojadong 192-1, 200-701, Chuncheon, Gangwon</wicri:regionArea><wicri:noRegion>Gangwon</wicri:noRegion></affiliation></author><author><name sortKey="Han, Seon Sook" sort="Han, Seon Sook" uniqKey="Han S" first="Seon-Sook" last="Han">Seon-Sook Han</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Internal Medicine, School of Medicine, Kangwon National University, Hyojadong 192-1, 200-701, Chuncheon, Gangwon</wicri:regionArea><wicri:noRegion>Gangwon</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Se Ran" sort="Yang, Se Ran" uniqKey="Yang S" first="Se-Ran" last="Yang">Se-Ran Yang</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Thoracic and Cardiovascular Surgery, School of Medicine, Kangwon National University, Chuncheon</wicri:regionArea><wicri:noRegion>Chuncheon</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Stem Cell Institute, Kangwon National University, Chuncheon</wicri:regionArea><wicri:noRegion>Chuncheon</wicri:noRegion></affiliation></author><author><name sortKey="Woo, Heung Myong" sort="Woo, Heung Myong" uniqKey="Woo H" first="Heung-Myong" last="Woo">Heung-Myong Woo</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Stem Cell Institute, Kangwon National University, Chuncheon</wicri:regionArea><wicri:noRegion>Chuncheon</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>College of Veterinary Medicine, Kangwon National University, Chuncheon</wicri:regionArea><wicri:noRegion>Chuncheon</wicri:noRegion></affiliation></author><author><name sortKey="Park, Sung Min" sort="Park, Sung Min" uniqKey="Park S" first="Sung-Min" last="Park">Sung-Min Park</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Thoracic and Cardiovascular Surgery, School of Medicine, Kangwon National University, Chuncheon</wicri:regionArea><wicri:noRegion>Chuncheon</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Stem Cell Institute, Kangwon National University, Chuncheon</wicri:regionArea><wicri:noRegion>Chuncheon</wicri:noRegion></affiliation></author><author><name sortKey="Lee, Hyang Ah" sort="Lee, Hyang Ah" uniqKey="Lee H" first="Hyang-Ah" last="Lee">Hyang-Ah Lee</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Obstetrics and Gynecology, School of Medicine, Kangwon National University, Hyojadong 192-1, 200-701, Chuncheon, Gangwon</wicri:regionArea><wicri:noRegion>Gangwon</wicri:noRegion></affiliation><affiliation></affiliation></author><author><name sortKey="Hong, Seok Ho" sort="Hong, Seok Ho" uniqKey="Hong S" first="Seok-Ho" last="Hong">Seok-Ho Hong</name><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Department of Internal Medicine, School of Medicine, Kangwon National University, Hyojadong 192-1, 200-701, Chuncheon, Gangwon</wicri:regionArea><wicri:noRegion>Gangwon</wicri:noRegion></affiliation><affiliation wicri:level="1"><country xml:lang="fr">Corée du Sud</country><wicri:regionArea>Stem Cell Institute, Kangwon National University, Chuncheon</wicri:regionArea><wicri:noRegion>Chuncheon</wicri:noRegion></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Corée du Sud</country></affiliation><affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">In Vitro Cellular & Developmental Biology - Animal</title><title level="j" type="abbrev">In Vitro Cell.Dev.Biol.-Animal</title><idno type="ISSN">1071-2690</idno><idno type="eISSN">1543-706X</idno><imprint><publisher>Springer-Verlag</publisher><pubPlace>New York</pubPlace><date type="published" when="2013-06-01">2013-06-01</date><biblScope unit="volume">49</biblScope><biblScope unit="issue">6</biblScope><biblScope unit="page" from="458">458</biblScope><biblScope unit="page" to="464">464</biblScope></imprint><idno type="ISSN">1071-2690</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1071-2690</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>GeneFishing</term><term>MEFs</term><term>SPARC</term><term>hPSC</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Mitotically inactivated feeder cells such as mouse embryonic fibroblast (MEFs) cells have been widely applied for physical and physiological support in the pluripotency maintenance of human pluripotent stem cells (hPSCs). However, accurate supporting mechanism or factors of feeder cells are poorly understood. Here, we isolated differentially expressed genes between wild-type MEFs and mitotically inactivated MEFs (miMEFs) by employing annealing control primer-based GeneFishing polymerase chain reaction. We identified a secreted protein acidic cysteine-rich glycoprotein (SPARC) gene that is upregulated in miMEFs. Suppression of SPARC expression in miMEFs using small interference RNA (siRNA) displayed gradual detachment of miMEFs. Furthermore, we found a significant reduction of OCT4- and SSEA3-positive hPS cell population maintained on SPARC siRNA-miMEFs compared to on miMEFs by flow cytometrical analysis. These findings suggest that SPARC plays a critical role in the maintenance of miMEFs without loss of cell number and might be a key component for supporting the culture of hPSCs.</div></front></TEI><affiliations><list><country><li>Corée du Sud</li><li>États-Unis</li></country></list><tree><country name="Corée du Sud"><noRegion><name sortKey="Won, Jun Yeon" sort="Won, Jun Yeon" uniqKey="Won J" first="Jun Yeon" last="Won">Jun Yeon Won</name></noRegion><name sortKey="Han, Seon Sook" sort="Han, Seon Sook" uniqKey="Han S" first="Seon-Sook" last="Han">Seon-Sook Han</name><name sortKey="Hong, Seok Ho" sort="Hong, Seok Ho" uniqKey="Hong S" first="Seok-Ho" last="Hong">Seok-Ho Hong</name><name sortKey="Hong, Seok Ho" sort="Hong, Seok Ho" uniqKey="Hong S" first="Seok-Ho" last="Hong">Seok-Ho Hong</name><name sortKey="Hong, Seok Ho" sort="Hong, Seok Ho" uniqKey="Hong S" first="Seok-Ho" last="Hong">Seok-Ho Hong</name><name sortKey="Kim, Woo Jin" sort="Kim, Woo Jin" uniqKey="Kim W" first="Woo Jin" last="Kim">Woo Jin Kim</name><name sortKey="Lee, Hyang Ah" sort="Lee, Hyang Ah" uniqKey="Lee H" first="Hyang-Ah" last="Lee">Hyang-Ah Lee</name><name sortKey="Lee, Seung Joon" sort="Lee, Seung Joon" uniqKey="Lee S" first="Seung-Joon" last="Lee">Seung-Joon Lee</name><name sortKey="Park, Sung Min" sort="Park, Sung Min" uniqKey="Park S" first="Sung-Min" last="Park">Sung-Min Park</name><name sortKey="Park, Sung Min" sort="Park, Sung Min" uniqKey="Park S" first="Sung-Min" last="Park">Sung-Min Park</name><name sortKey="Woo, Heung Myong" sort="Woo, Heung Myong" uniqKey="Woo H" first="Heung-Myong" last="Woo">Heung-Myong Woo</name><name sortKey="Woo, Heung Myong" sort="Woo, Heung Myong" uniqKey="Woo H" first="Heung-Myong" last="Woo">Heung-Myong Woo</name><name sortKey="Yang, Se Ran" sort="Yang, Se Ran" uniqKey="Yang S" first="Se-Ran" last="Yang">Se-Ran Yang</name><name sortKey="Yang, Se Ran" sort="Yang, Se Ran" uniqKey="Yang S" first="Se-Ran" last="Yang">Se-Ran Yang</name></country><country name="États-Unis"><noRegion><name sortKey="Lee, Young Jin" sort="Lee, Young Jin" uniqKey="Lee Y" first="Young Jin" last="Lee">Young Jin Lee</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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