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Lee, Youn-Su,Ock, Sun-A,Cho, Seong-Keun,Jeon, Byeong-Gyun,Kang, Tae-Young,Balasubramanian, S.,Choe, Sang-Yong,Rho, Gyu-Jin Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.5
In this study, two cell types from porcine females, namely fetal fibroblasts (pFFs) and adult ear fibroblasts (pAEFs) and two passages (3-4 and 7-8) were investigated by evaluating the development rate, blastocyst cell number and the incidence of apoptosis. No significant differences were observed in the cleavage rates of cloned and IVF embryos. The blastocyst rates between the embryos cloned with pFFs ($15.1{\pm}3.2$) and pAEFs ($10.4{\pm}2.6$) did not differ significantly but was significantly (p<0.05) lower in pAEFs than that in IVF ($22.5{\pm}4.5$) embryos. Total cell number in pFFs ($28.4{\pm}4.3$) and pAEFs cloned blastocysts ($24.2{\pm}5.1$) was significantly (p<0.05) lesser than IVF control ($35.4{\pm}3.2$). Apoptosis rates between cloned blastocysts differed significantly (p<0.05) and were significantly (p<0.05) higher than IVF embryos. The blastocyst rates between the cloned embryos cloned with different cell passages did not differ significantly but in embryos cloned with 7-8 cell passage was significantly (p<0.05) lower than the IVF control. Apoptosis signals were detected in IVF and cloned embryos as early as day 3 and the rates of apoptosis increased concurrently with the embryo development. In conclusion, high apoptosis during in vitro preimplantation development resulted in low development rate and total cell number of cloned embryos. Moreover, based on the apoptotic incidence in cloned blastocysts, fetal fibroblasts are more suitable for production of cloned embryos in porcine.
Jeong, Young Sun,Oh, Keon Bong,Park, Jung Sun,Kim, Ji-Su,Kang, Yong-Kook Wiley-Liss, Inc. 2009 Developmental dynamics Vol.238 No.7
<P>DNA methyltransferase-1 (Dnmt1) is involved in the maintenance of genomic methylation patterns. Rather than full-length Dnmt1, mouse oocytes have a truncated variant called Dnmt1o. Immunofluorescence data showed that Dnmt1o localized to the cytoplasm, but this has not been confirmed using more direct methods. The cytoplasmic localization of Dnmt1o has been assigned to the main cause of global DNA demethylation in early mouse embryos. We studied localization of Dnmt1o in mouse and pig embryos. We identified pig Dnmt1o protein and its transcript with unique 5′-end sequence. Physically separating mouse and pig 2-cell embryos into their nuclear and cytoplasmic components demonstrated that Dnmt1o of both species localized to the cytoplasm. Cloned pig embryos had Dnmt1o as the main form, with no indication of somatic Dnmt1. These findings indicate that Dnmt1o is cytoplasmic during early development; its presence in both pig and mouse embryos further suggests that Dnmt1o is conserved in mammals. Developmental Dynamics 238:1666–1673, 2009. © 2009 Wiley-Liss, Inc.</P>
권대진,오건봉,옥선아,이정웅,이승수,박진기,장원경,황성수 사단법인 한국동물생명공학회 2013 Reproductive & developmental biology Vol.36 No.4
This study aimed at investigating whether a porcine follicular fluid (pFF) supplementation positively affects the characteristics of donor cells and the developmental competence of porcine cloned embryos. Ear fibroblast cells (donor cell) from an Massachusetts General Hospital miniature pig were cultured in different culture methods: (1) Dulbecco's modified Eagle's medium (DMEM)+10% FBS (Control); (2) DMEM+0.5% FBS (SS); and (3) DMEM+10% FBS+10% pFF (pFF) for 72 h. In each conditioned medium, the concentrations of 4 amino acids (Thr, Glu, Pro, and Val) in the pFF group were significantly different from those in the control group (p<0.05 or p<0.01). The proliferation of the cells cultured in the SS group was significantly lower than that of the other treatment groups (p<0.01). The population of apoptotic and necrotic cells in the SS group was significantly higher than that of either the control or the pFF group (p<0.01). The number of embryos that cleaved (p<0.05) and developed into blastocysts (p<0.01) in the SS group was significantly lower than that of either the control or the pFF group. Compared to other groups, the blastocysts produced from the donor cells in the pFF group had higher total cells and lower apoptotic cells (p<0.05). It can be concluded that pFF supplementation in the donor cell culture medium positively affects cell death, cell cycle and quality of the cloned blastocyst.
Kwon, Dae-Jin,Oh, Keon Bong,Ock, Sun A,Lee, Jeong Woong,Lee, Sung-Soo,Park, Jin-Ki,Chang, Won-Kyong,Hwang, Seongsoo The Korean Society of Animal Reproduction 2012 Reproductive & developmental biology Vol.36 No.4
This study aimed at investigating whether a porcine follicular fluid (pFF) supplementation positively affects the characteristics of donor cells and the developmental competence of porcine cloned embryos. Ear fibroblast cells (donor cell) from an Massachusetts General Hospital miniature pig were cultured in different culture methods: (1) Dulbecco's modified Eagle's medium (DMEM)+10% FBS (Control); (2) DMEM+0.5% FBS (SS); and (3) DMEM+10% FBS+10% pFF (pFF) for 72 h. In each conditioned medium, the concentrations of 4 amino acids (Thr, Glu, Pro, and Val) in the pFF group were significantly different from those in the control group (p<0.05 or p<0.01). The proliferation of the cells cultured in the SS group was significantly lower than that of the other treatment groups (p<0.01). The population of apoptotic and necrotic cells in the SS group was significantly higher than that of either the control or the pFF group (p<0.01). The number of embryos that cleaved (p<0.05) and developed into blastocysts (p<0.01) in the SS group was significantly lower than that of either the control or the pFF group. Compared to other groups, the blastocysts produced from the donor cells in the pFF group had higher total cells and lower apoptotic cells (p<0.05). It can be concluded that pFF supplementation in the donor cell culture medium positively affects cell death, cell cycle and quality of the cloned blastocyst.
Developmental Efficiency of Bovine SCNT Embryos from the Cells with Arhgap15 Gene Knock-Down
Byoung-Chul Yang,Hwi-Cheul Lee,Seongsoo Hwang,Jae-Seok Woo,Ik-Soo Jeon,Dong-Kyeong Lee,Na-Yeong Gu,Jin-Hee An,Young-Joon Han,Eun-Young Kim,Soo-Bong Park 한국동물번식학회 2010 Reproductive & Developmental Biology(Supplement) Vol.34 No.2s
Developmental Efficiency of Bovine SCNT Embryos from the Cells with Arhgap15 Gene Knock-Down
Byoung-Chul Yang,Hwi-Cheul Lee,Seongsoo Hwang,Jae-Seok Woo,Ik-Soo Jeon,Dong-Kyeong Lee,Na-Yeong Gu,Jin-Hee An,Young-Joon Han,Eun-Young Kim,Soo-Bong Park 한국동물생명공학회(구 한국동물번식학회) 2010 Reproductive & developmental biology Vol.34 No.2