title: Cell Synchronization by Rapamycin Improves the Developmental Competence of Porcine SCNT Embryos.
abstract: The cell cycle stage of donor cells influences the success of somatic cell nuclear transfer (SCNT). This study investigated the effects of rapamycin treatment on synchronization of porcine fibroblasts in comparison with control and serum-starved cells, SCNT donor cell viability, and SCNT-derived embryo development. Porcine fibroblasts were treated with 0.1, 1, 10, and 100 μM rapamycin for 1 or 3 days. The proportion of cells in G0/G1 phase was significantly higher among cells treated with 1 μM rapamycin for 3 days (D3-1R) than among control and serum-starved cells (p < 0.05). In comparison with control cells, rapamycin-treated cells exhibited reduced proliferation, similar to serum-starved cells. The viability (as assessed by the MTT assay) of D3-1R-treated cells was good, similar to control cells, showing their quality was maintained. To confirm nutrient regulation by rapamycin treatment, we checked the transcript levels of nutrient transporter genes (SLC2A2, SLC2A4, SLC6A14, and SLC7A1). These levels were significantly lower in D3-1R-treated cells than in control cells (p < 0.01). We performed SCNT with D3-1R-treated cells (SCNT(D3-1R)) to confirm the effect of cell cycle synchronization by rapamycin treatment. Although SCNT(D3-1R) embryos did not have an increased fusion rate, their cleavage and blastocyst formation rates were significantly higher than those of control embryos (p < 0.05). Regarding embryo quality, the numbers of total and apoptotic cells per blastocyst were increased and decreased, respectively, in SCNT(D3-1R) blastocysts. The mRNA levels of developmental (CDX2 and CDH1) and proapoptotic (FAS and CASP3) genes were significantly higher and lower, respectively, in SCNT(D3-1R) blastocysts than in control blastocysts (p < 0.05). These results demonstrate that rapamycin treatment affects the cell cycle synchronization of donor cells and enhances the developmental potential of porcine SCNT embryos.
authors.affiliation: 1 Stem cell Research Center, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.
authors.forename: Hyuk
authors.initials: H
authors.lastname: Hyun
authors.affiliation: 1 Stem cell Research Center, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.
authors.forename: Seung-Eun
authors.initials: SE
authors.lastname: Lee
authors.affiliation: 1 Stem cell Research Center, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.
authors.forename: Yeo-Jin
authors.initials: YJ
authors.lastname: Son
authors.affiliation: 1 Stem cell Research Center, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.
authors.forename: Min-Young
authors.initials: MY
authors.lastname: Shin
authors.affiliation: 1 Stem cell Research Center, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.
authors.forename: Yun-Gwi
authors.initials: YG
authors.lastname: Park
authors.affiliation: 1 Stem cell Research Center, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|3 Mirae Cell Bio , Gwangjin-gu, Seoul, Korea.
authors.forename: Eun-Young
authors.initials: EY
authors.lastname: Kim
authors.affiliation: 1 Stem cell Research Center, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|2 Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University , Jeju, Jeju Special Self-Governing Province, Korea.|3 Mirae Cell Bio , Gwangjin-gu, Seoul, Korea.
authors.forename: Se-Pill
authors.initials: SP
authors.lastname: Park
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects*; D000071616:CDX2 Transcription Factor / Q000235:genetics / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D005307:Fertilization in Vitro; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects*; D018507:Gene Expression Regulation, Developmental; D007399:Interphase; D053652:Nuclear Transfer Techniques; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020123:Sirolimus / Q000494:pharmacology*; D013552:Swine
chemical_list: D000071616:CDX2 Transcription Factor; D012333:RNA, Messenger; D020123:Sirolimus