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Cell Journal، جلد ۱۹، شماره ۴، صفحات ۶۴۰-۶۴۶
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عنوان انگلیسی Effects of Klf4 and c-Myc Knockdown on Pluripotency Maintenance in Porcine Induced Pluripotent Stem Cell
چکیده انگلیسی مقاله Objective: The importance of Oct4 and Sox2 in maintaining pluripotency and self-renewal is well-understood, but the function of Klf4 and c-Myc has not been fully investigated. In the present study, we attempted to determine the roles of Klf4 and c-Myc on pluripotency maintenance of porcine induced pluripotent stem cells (piPS cells). Materials and methods: In this study, we performed short hairpin RNA (shRNA) to knock down the Klf4 and c-Myc function of piPS cells and examined pluripotency markers and teratoma formation to evaluate piPS cell pluripotency. The shRNA-Klf4 and shRNA-c-Myc vectors containing a reporter gene, TagFP635, were transfected into piPS cells by lentivirus infection. The piPS cells fully expressing infrared fluorescence were selected to confirm gene knockdown of Klf4 and c-Myc by reverse-transcription polymerase chain reaction (RT-PCR). Next, for pluripotency evaluation, expression of pluripotency markers was detected by immunocytochemical staining, and capability of teratoma formation was investigated by piPS cell transplantation into NOD-SCID mice. Results: Our findings indicated that Klf4 and c-Myc function of piPS cells were knocked down by shRNA transfection, and knockdown of Klf4 and c-Myc function impaired expression of pluripotency markers such as Oct4, AP, SSEA-3, SSEA-4, TRA-1-6, and TRA-1-81. Furthermore, piPS cells without Klf4 and c-Myc expression failed to form teratomas. Conclusion: The pluripotency of piPS cells are crucially dependent upon Klf4 and c-Myc expression. These findings, suggesting potential mechanisms of Klf4 and c-Myc contribution to piPS cell formation, have important implications for application, regulation, and tumorigenesis of piPS cells.
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نویسندگان مقاله | Yu-Jing Liao


| Yi-Shiou Chen


| Ja-Xin Lee


| Lih-Ren Chen


| Jenn-Rong Yang


نشانی اینترنتی http://celljournal.org/journal/article/abstract/4428
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