title: SPARC is involved in the maintenance of mitotically inactivated mouse embryonic fibroblast cells.
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.
authors: Department of Otolaryngology, School of Medicine, Kangwon National University, Chuncheon, South Korea.
authors: Jun Yeon
authors: JY
authors: Won
authors: Young Jin
authors: YJ
authors: Lee
authors: Seung-Joon
authors: SJ
authors: Lee
authors: Woo Jin
authors: WJ
authors: Kim
authors: Seon-Sook
authors: SS
authors: Han
authors: Se-Ran
authors: SR
authors: Yang
authors: Heung-Myong
authors: HM
authors: Woo
authors: Sung-Min
authors: SM
authors: Park
authors: Hyang-Ah
authors: HA
authors: Lee
authors: Seok-Ho
authors: SH
authors: Hong
mesh_terms: D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D017077:Culture Media, Conditioned; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D051379:Mice; D008938:Mitosis / Q000235:genetics*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D015676:Osteonectin / Q000235:genetics* / Q000378:metabolism; D055746:Stage-Specific Embryonic Antigens / Q000378:metabolism
chemical_list: D015295:Antigens, Tumor-Associated, Carbohydrate; D017077:Culture Media, Conditioned; D050814:Octamer Transcription Factor-3; D015676:Osteonectin; C494174:Pou5f1 protein, mouse; C578250:SPARC protein, mouse; D055746:Stage-Specific Embryonic Antigens; C052924:stage-specific embryonic antigen-3
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