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embryonic development following somatic cell nuclear transfer

Hanna Lee, Fazle Elahi, Joohyeong Lee, Seung Tae Lee, Sang-Hwan Hyun, Eunsong Lee
This study was conducted to evaluate the effects of cyclic AMP (cAMP) modulator cilostamide (CIL) and forskolin (FSK) treatment during in vitro growth (IVG) on growth, maturation, and embryonic development of cumulus-oocyte complexes (COCs) derived from small antral follicles < 3 mm in diameter (SAFCOCs). SAFCOCs were untreated (control) or treated with 20 μM CIL and/or 50 μM FSK for 2 days for IVG. Next, IVG oocytes were cultured for maturation and then induced for parthenogenesis (PA) or used as recipient ooplasts for somatic cell nuclear transfer (SCNT)...
March 15, 2017: Theriogenology
F Moulavi, S M Hosseini, N Tanhaie-Vash, S Ostadhosseini, S H Hosseini, M Hajinasrollah, M H Asghari, H Gourabi, A Shahverdi, A D Vosough, M H Nasr-Esfahani
Recent accomplishments in the field of somatic cell nuclear transfer (SCNT) hold tremendous promise to prevent rapid loss of animal genetic resources using ex situ conservation technology. Most of SCNT studies use viable cells for nuclear transfer into recipient oocytes. However, preparation of live cells in extreme circumstances, in which post-mortem material of endangered/rare animals is improperly retained frozen, is difficult, if not impossible. This study investigated the possibility of interspecies-SCNT (iSCNT) in Asiatic cheetah (Acinonyx jubatus venaticus), a critically endangered subspecies, using nuclei derived from frozen tissue in absence of cryo-protectant at -20 °C and in vitro matured domestic cat oocytes...
March 1, 2017: Theriogenology
H L Sun, L N Meng, X Zhao, J R Jiang, Q Y Liu, D S Shi, F H Lu
Buffalo are characteristic livestock of the Guangxi Zhuang Autonomous Region of China, but their low reproductive capacity necessitates the use of somatic cell nuclear transfer (SCNT). We investigated the effects of RG108 on DNA methylation in buffalo adult fibroblasts, and on subsequent SCNT embryo development. RG108 treatment (0, 5, 10, 20, and 100 mM) had no effect on cell morphology, viability, or karyotype (2n = 48), and cell growth followed a typical "S" curve. Immunohistochemistry showed that relative DNA methylation gradually decreased as RG108 concentration increased, and was significantly lower in the 20 and 100 mM groups compared to the 0, 5, and 10 mM treatments (0...
September 2, 2016: Genetics and Molecular Research: GMR
Min Yang, Justin Hall, Zhiqiang Fan, Misha Regouski, Qinggang Meng, Heloisa M Rutigliano, Rusty Stott, Kerry A Rood, Kip E Panter, Irina A Polejaeva
Reduced developmental competence after IVF has been reported using oocyte derived from small follicles in several species including cattle, sheep, and goats. No information is currently available about the effect of follicle size of the cytoplast donor on in vivo development after somatic cell nuclear transfer (SCNT) in goats. Oocytes collected from large (≥3 mm) and small follicles (<3 mm) were examined for maturation and in vivo developmental competence after SCNT. Significantly greater maturation rate was observed in oocytes derived from large follicles compared with that of small follicles (51...
December 2016: Theriogenology
Heiner Niemann
The birth of "Dolly," the first mammal cloned from an adult mammary epithelial cell, abolished the decades-old scientific dogma implying that a terminally differentiated cell cannot be reprogrammed into a pluripotent embryonic state. The most dramatic epigenetic reprogramming occurs in SCNT when the expression profile of a differentiated cell is abolished and a new embryo-specific expression profile, involving 10,000 to 12,000 genes, and thus, most genes of the entire genome is established, which drives embryonic and fetal development...
July 1, 2016: Theriogenology
Jan O Secher, Henrik Callesen, Kristine K Freude, Poul Hyttel
The quest for porcine pluripotent stem cells (PSCs) was initiated in the early 90s. Initially, it was the intention to benefit from these cells for production of genetically modified pigs using homologous recombination followed by derivation of chimeric offspring; a technology that has been used to produce genetically modified mice since the mid-80s. However, no convincing reports on the generation of bona fide porcine embryonic stem cells or embryonic germ cells resulted from these activities, and with the advent of somatic cell nuclear transfer during the late 90s, alternative methods for creating genetically modified pigs emerged...
January 1, 2016: Theriogenology
Joohyeong Lee, Jong-Im Park, Geun-Shik Lee, Jung Hoon Choi, Seung Tae Lee, Choon-Keun Park, Dae Young Kim, Sang-Hwan Hyun, Eunsong Lee
The objective of this study was to examine the effects of colcemid treatment during oocyte in vitro maturation (IVM) and embryonic development after parthenogenetic activation (PA) and somatic-cell nucleus transfer (SCNT) in pigs. Immature oocytes were treated with colcemid from 0 to 22, 38 to 42, or 0 to 22 hr followed by 38 to 42 hr during IVM (designated as COL0-22, COL38-42, and COL0-22/38-42, respectively). The proportion of oocytes reaching the germinal vesicle (GV)/GV breakdown (GVBD) stage after 22 hr of IVM was higher in COL0-22 (98...
June 2015: Molecular Reproduction and Development
S H Lee, E J Park, J H Moon, K Y Song, S J Kim, J K Cho, B C Lee
Antioxidants are widely used for in vitro production of embryos due to their activity as reactive oxygen species scavengers. Among various antioxidants, resveratrol supplementation in in vitro-maturation (IVM) media and trolox supplementation in in vitro-culture (IVC) media improves oocyte maturation and embryonic development in other species, such as cattle and sheep. Limited information is available, however, on the effect of resveratrol and/or trolox on porcine embryos produced in vitro. In this study, we evaluated the effect of resveratrol supplemented to the media of IVM and trolox treatment during IVC on porcine parthenotes...
December 2014: Reproduction, Fertility, and Development
L T K Do, Y Sato, M Taniguchi, T Otoi
The developmental ability of interspecies somatic cell nuclear transfer (iSCNT) embryos decreases as the taxonomic distance between the donor and recipient species increases. Treatment of cat iSCNT embryos using bovine oocytes with 50nM of trichostatin A (TSA) improves in vitro embryonic development (Wittayarat et al. 2013 Cell. Reprogram. 15, 301-308). This study investigated whether the TSA treatment effects differ between the development of cat iSCNT embryos reconstructed with porcine and bovine oocytes...
December 2014: Reproduction, Fertility, and Development
Shogo Matoba, Yuting Liu, Falong Lu, Kumiko A Iwabuchi, Li Shen, Azusa Inoue, Yi Zhang
Mammalian oocytes can reprogram somatic cells into a totipotent state enabling animal cloning through somatic cell nuclear transfer (SCNT). However, the majority of SCNT embryos fail to develop to term due to undefined reprogramming defects. Here, we identify histone H3 lysine 9 trimethylation (H3K9me3) of donor cell genome as a major barrier for efficient reprogramming by SCNT. Comparative transcriptome analysis identified reprogramming resistant regions (RRRs) that are expressed normally at 2-cell mouse embryos generated by in vitro fertilization (IVF) but not SCNT...
November 6, 2014: Cell
Hiren Patel, Shruti Chougule, Parul Chohan, Naval Shah, Deepa Bhartiya
Pluripotent stem cells are possibly the best candidates for regenerative medicine, and somatic cell nuclear transfer (SCNT) is one of the viable options to make patient-specific embryonic stem cells. Till date efficacy of SCNT embryos is very low and requires further improvement like ideal oocyte activation and in vitro culture system. The aim of the present study was to evaluate ideal oocyte activation using different stimulation protocols and to study the effect of cumulus co-culture conditions on embryo development...
October 2014: Indian Journal of Experimental Biology
Naru Zhou, Zubing Cao, Ronghua Wu, Xing Liu, Jia Tao, Zhen Chen, Dandan Song, Fei Han, Yunsheng Li, Fugui Fang, Xiaorong Zhang, Yunhai Zhang
Histone H3 lysine 27 acetylation (H3K27ac) is an active epigenetic modification which has been revealed to be associated with active gene expression. It was hypothesized that H3K27ac might also participate in the porcine somatic reprogramming process during early development of SCNT-derived embryos. The spatial and temporal expression profiles of H3K27ac were investigated at different developmental stages in SCNT embryos compared with in vitro fertilization (IVF) and parthenogenetic activation (PA) counterparts...
August 2014: Animal Reproduction Science
Gareth D Greggains, Lisa M Lister, Helen A L Tuppen, Qi Zhang, Louise H Needham, Nilendran Prathalingam, Louise A Hyslop, Lyndsey Craven, Zbigniew Polanski, Alison P Murdoch, Douglass M Turnbull, Mary Herbert
Induced pluripotent stem cells (iPSCs) hold much promise in the quest for personalised cell therapies. However, the persistence of founder cell mitochondrial DNA (mtDNA) mutations limits the potential of iPSCs in the development of treatments for mtDNA disease. This problem may be overcome by using oocytes containing healthy mtDNA, to induce somatic cell nuclear reprogramming. However, the extent to which somatic cell mtDNA persists following fusion with human oocytes is unknown. Here we show that human nuclear transfer (NT) embryos contain very low levels of somatic cell mtDNA...
January 24, 2014: Scientific Reports
Geon A Kim, Hyun Ju Oh, Tae Hee Lee, Ji Hyun Lee, Sang Hwan Oh, Ju Hyun Lee, Jin Wook Kim, Se Woon Kim, Byeong Chun Lee
Canine adipose-derived mesenchymal stem cells (ASCs) are promising as donor cells for somatic cell nuclear transfer (SCNT). It has been suggested that different cell cultures possess different capacities to support pre-implantation development of SCNT embryos. The aim of this study is to investigate whether two culture medium (RCMEP, Dulbecco's modified Eagle's medium [DMEM]) affect gene expression of ASCs, subsequent development of interspecies SCNT (iSCNT) and gene expression of cloned embryos. The RCMEP-cultured cells contained significantly greater amounts of SOX2, NANOG, OCT4, DNMT1, and MeCP2 than DMEM-cultured cells (P < 0...
January 15, 2014: Theriogenology
Chih-Jie Shen, Chiao-Chieh Lin, Perng-Chih Shen, Winston T K Cheng, Hsiao-Ling Chen, Tsung-Chou Chang, Shyh-Shyan Liu, Chuan-Mu Chen
In mammals, genome-wide epigenetic reprogramming systems exist in primordial germ cells and zygotes. These reprogramming systems play crucial roles in regulating genome functions during critical stages of embryonic development, and they confer the stability of gene expression during mammalian development. The frequent unexpected loss of progeny from somatic cell nuclear transfer (SCNT) is an ongoing problem. In this study, we used six cloned bovines (named NT-1 to NT-6), which were created by ear fibroblast nuclear transfer and displayed short life spans with multiple organ defects, as an experimental model...
October 2013: Cellular Reprogramming
V Hall, K Hinrichs, G Lazzari, D H Betts, P Hyttel
Over many decades assisted reproductive technologies, including artificial insemination, embryo transfer, in vitro production (IVP) of embryos, cloning by somatic cell nuclear transfer (SCNT), and stem cell culture, have been developed with the aim of refining breeding strategies for improved production and health in animal husbandry. More recently, biomedical applications of these technologies, in particular, SCNT and stem cell culture, have been pursued in domestic mammals in order to create models for human disease and therapy...
August 2013: Veterinary Journal
Manita Wittayarat, Yoko Sato, Lanh Thi Kim Do, Yasuhiro Morita, Kaywalee Chatdarong, Mongkol Techakumphu, Masayasu Taniguchi, Takeshige Otoi
Abnormal epigenetic reprogramming, such as histone acetylation, might cause low efficiency of interspecies somatic cell nuclear transfer (iSCNT). This study was conducted to evaluate the effects of trichostatin A (TSA) on the developmental competence and histone acetylation of iSCNT embryos reconstructed from cat somatic cells and bovine cytoplasm. The iSCNT cat and parthenogenetic bovine embryos were treated with various concentrations of TSA (0, 25, 50, or 100 nM) for 24 h, respectively, following fusion and activation...
August 2013: Cellular Reprogramming
Jae-Il Bang, Jae-Gyu Yoo, Mi-Rung Park, Teak-Soon Shin, Byung-Wook Cho, Hong-Gu Lee, Byeong-Woo Kim, Tae-Young Kang, Il-Keun Kong, Jin-Hoi Kim, Seong-Keun Cho
This study investigated the effects of two different activation regimens on the developmental potential of somatic cell nuclear transfer (SCNT) embryos and postnatal survivability of the cloned piglets. In vitro matured oocytes were enucleated and reconstructed with porcine fetal fibroblasts. On the basis of the activation regimen used, the reconstructed porcine embryos were allocated into two groups: Group 1-simultaneous electrical pulses and activation group (SFA group); and Group 2-electrical fusion without calcium followed by electrical pulses with calcium after colcemid and cytochalasin B treatment for 5h (DA group)...
June 2013: Reproductive Biology
Jun Liu, Zhongcai Yang, Mingning Qiu, Yan Luo, Meijun Pang, Yongyan Wu, Yong Zhang
Previous studies have demonstrated that skin stem cells expressing the pluripotency marker stage-specific embryonic antigen 3 (SSEA3) are easier to reprogram into induced pluripotent stem cells (iPSCs) than skin fibroblasts. Furthermore, it is widely speculated that the undifferentiated state may make stem cells more efficient donor cells for somatic cell nuclear transfer (SCNT). In this study, we isolated SSEA3(+) cells from goat skin fibroblast cells (SFCs) using fluorescence-activated cell sorting (FACS) and examined expression of pluripotency markers and in vitro development of cloned embryos following SCNT...
April 2013: Cellular Reprogramming
Yong-Nan Xu, Sang-Jun Uhm, Bon-Chul Koo, Mo-Sun Kwon, Ji-Yeol Roh, Jung-Seok Yang, Hyun-Yong Choi, Young-Tae Heo, Xiang-Shun Cui, Joon-Ho Yoon, Dae-Hwan Ko, Teoan Kim, Nam-Hyung Kim
The potential benefits of generating and using transgenic cattle range from improvements in agriculture to the production of large quantities of pharmaceutically relevant proteins. Previous studies have attempted to produce transgenic cattle and other livestock by pronuclear injection and somatic cell nuclear transfer, but these approaches have been largely ineffective; however, a third approach, lentivirus-mediated transgenesis, has successfully produced transgenic livestock. In this study, we generated transgenic (TG) Korean native cattle using perivitelline space injection of viral vectors, which expressed enhanced green fluorescent protein (EGFP) systemically...
January 20, 2013: Journal of Genetics and Genomics, Yi Chuan Xue Bao
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