Cloning Stem Cells最新文献_第5页

Improved efficacy and safety of in utero cell transplantation in sheep using an ultrasound-guided method. 超声引导下绵羊子宫内细胞移植的疗效和安全性提高。

Cloning Stem Cells Pub Date : 2009-06-01 DOI: 10.1089/clo.2008.0067

Yoshikazu Nagao, Tomoyuki Abe, Hideaki Hasegawa, Yujiro Tanaka, Kyoko Sasaki, Yoshihiro Kitano, Satoshi Hayashi, Yutaka Hanazono

{"title":"Improved efficacy and safety of in utero cell transplantation in sheep using an ultrasound-guided method.","authors":"Yoshikazu Nagao, Tomoyuki Abe, Hideaki Hasegawa, Yujiro Tanaka, Kyoko Sasaki, Yoshihiro Kitano, Satoshi Hayashi, Yutaka Hanazono","doi":"10.1089/clo.2008.0067","DOIUrl":"https://doi.org/10.1089/clo.2008.0067","url":null,"abstract":"In the present study, we investigated the suitability of two methods for the transplantation of cells into ovine fetuses. The first method was an ultrasound-guided cell injection via the uterine wall. The second involved hysterotomic cell injection with an incision in the uterine wall exposing the amnion. Monkey embryonic stem (ES) cell-derived hematopoietic cells were used as donor cells. After transplantation, the abortion rate associated with the hysterotomic injection method was significantly higher than that of the ultrasound-guided injection method (8/13 versus 4/24; P < 0.01). The fetuses were delivered to examine the engraftment of transplanted monkey hematopoietic cells. Monkey cells were detected in one of the five animals (20%) in the hysterotomic injection group, and 14 of 20 animals (70%, P < 0.05) in the ultrasound-guided injection group. Therefore, the ultrasound-guided method was effectively shown to be minimally invasive for in utero transplantation and can produce a higher rate of engraftment for transplanted cells.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 2","pages":"281-5"},"PeriodicalIF":0.0,"publicationDate":"2009-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0067","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28314993","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 11

Reprogramming of active and repressive histone modifications following nuclear transfer with rabbit mesenchymal stem cells and adult fibroblasts. 兔间充质干细胞和成纤维细胞核移植后活性和抑制性组蛋白修饰的重编程。

Cloning Stem Cells Pub Date : 2009-06-01 DOI: 10.1089/clo.2008.0083

Alessandro Brero, Ru Hao, Matthias Schieker, Matthias Wierer, Eckhard Wolf, Thomas Cremer, Valeri Zakhartchenko

{"title":"Reprogramming of active and repressive histone modifications following nuclear transfer with rabbit mesenchymal stem cells and adult fibroblasts.","authors":"Alessandro Brero, Ru Hao, Matthias Schieker, Matthias Wierer, Eckhard Wolf, Thomas Cremer, Valeri Zakhartchenko","doi":"10.1089/clo.2008.0083","DOIUrl":"https://doi.org/10.1089/clo.2008.0083","url":null,"abstract":"Following nuclear transfer (NT) the epigenetic state of a donor nucleus must be reprogrammed to an embryonic one. To evaluate the efficiency of nuclear reprogramming, we monitored the levels of histone H3 di/tri-methylated on lysine 4 (H3K4m2/3), a marker for transcriptionally active/permissive euchromatin, and of histone H3 tri-methylated on lysine 27 (H3K27m3), a modification associated with facultative heterochromatin, in embryos cloned using rabbit mesenchymal stem cells (MSC) and adult fibroblasts (RAF) isolated from the same animals. In vivo fertilized, in vitro cultured embryos served as controls. H3K27m3 was undetectable in all stages of control embryos except for weak staining in a few blastocyst cells. A similar situation was found in all NT embryos irrespective of the type of donor cells used, although both MSC and RAF stained substantially for H3K27m3. H3K4m2/3 levels were very high in one- and two-cell control embryos, but then decreased to reach a minimum at the eight-cell stage, and finally increased again to initial levels at the morula and blastocyst stage. Reprogramming of H3K4m2/3 differed remarkably among the different types NT embryos as well as between NT embryos and control embryos, and was apparently dependent on the type of donor cells. Interestingly, abnormal reprogramming of H3K4m2/3 was observed in NT embryos derived from both MSC and RAF, donor cell types with markedly different proliferation capacity. Our study demonstrates that the repressive chromatin modification, H3K27m3, is faithfully reprogrammed in NT embryos derived from MSC or RAF, while reprogramming of the activating chromatin modification, H3K4m2/3, is quite variable and does not reflect the situation observed in control embryos derived by fertilization.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 2","pages":"319-29"},"PeriodicalIF":0.0,"publicationDate":"2009-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0083","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28231239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 13

Effect of sperm extract injection volume, injection of PLCzeta cRNA, and tissue cell line on efficiency of equine nuclear transfer. 精子提取物注射量、PLCzeta cRNA注射量和组织细胞系对马核移植效率的影响。

Cloning Stem Cells Pub Date : 2009-06-01 DOI: 10.1089/clo.2008.0077

Young-Ho Choi, David L Hartman, Rafael A Fissore, Sylvia J Bedford-Guaus, Katrin Hinrichs

{"title":"Effect of sperm extract injection volume, injection of PLCzeta cRNA, and tissue cell line on efficiency of equine nuclear transfer.","authors":"Young-Ho Choi, David L Hartman, Rafael A Fissore, Sylvia J Bedford-Guaus, Katrin Hinrichs","doi":"10.1089/clo.2008.0077","DOIUrl":"https://doi.org/10.1089/clo.2008.0077","url":null,"abstract":"We evaluated the effect of different activation methods on blastocyst development after equine nuclear transfer. All activation treatments were followed by incubation in 2 mM 6-dimethylaminopurine for 4 h. In Experiment 1, reconstructed oocytes were injected with sperm extract for 0.1, 0.2, 0.4, 0.8, or 1.6 sec using a FemtoJet injection device, then treated with ionomycin. The blastocyst rate (9.8%) for 0.1-sec injection was significantly higher than that for 0.2 sec (0%) or 0.8 sec (1.4%). In Experiment 2, injection of murine PLCzeta cRNA before or after ionomycin treatment did not increase blastocyst development (0 and 4.5%) over a control treatment (injection of sperm extract after ionomycin exposure; 5.6%). Transfer of 10 blastocysts produced in Experiments 1 and 2 resulted in five pregnancies, all lost before 70 days of gestation. In Experiment 3, cells from a second biopsy sample from the same horse produced significantly more blastocysts than did the original sample (4/44 vs. 0/58; p < 0.05). Transfer of these four blastocysts produced two viable foals. In Experiment 4, blastocyst development rates did not differ between oocytes in metaphase I or II at the time of nuclear transfer (16.7 and 3.0%, respectively). A healthy foal was produced from a blastocyst originating from a metaphase I oocyte.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 2","pages":"301-8"},"PeriodicalIF":0.0,"publicationDate":"2009-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0077","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28230073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 29

Efficiency of asynchronously in vitro-matured oocytes as recipients for nuclear transfer and of blind enucleation in zona-free bovine cloning. 非同步体外成熟卵母细胞作为核移植受体和牛无带克隆盲去核的效率。

Cloning Stem Cells Pub Date : 2009-06-01 DOI: 10.1089/clo.2007.0090

Galina P Malenko, Oleg I Stepanov, Andrey V Komissarov, Tatyana A Antipova, Marina V Pinyugina, Michael I Prokofiev

{"title":"Efficiency of asynchronously in vitro-matured oocytes as recipients for nuclear transfer and of blind enucleation in zona-free bovine cloning.","authors":"Galina P Malenko, Oleg I Stepanov, Andrey V Komissarov, Tatyana A Antipova, Marina V Pinyugina, Michael I Prokofiev","doi":"10.1089/clo.2007.0090","DOIUrl":"https://doi.org/10.1089/clo.2007.0090","url":null,"abstract":"We studied the dynamics of in vitro maturation of bovine oocytes, the efficiency of asynchronously matured oocytes as recipients for the generation of embryos produced by nuclear transfer, and the potential for using blind enucleation of zona-free bovine oocytes in bovine cloning. At 15 h after the initiation of maturation (hpm), oocytes were freed from both cumulus cells and the zona pellucida, and the dynamics of oocyte maturation were monitored every 30 min through the criterion of extrusion of the first polar body (PB1). More than 41% of bovine oocytes had extruded PB1 by 16.5 hpm, and were designated as representing a group of rapidly maturing oocytes. A second group, comprising about 25% of all oocytes, had extruded PB1 by 18.5-20.0 hpm. Examination of Hoechst 33342-stained samples demonstrated that PB1 on the surfaces of zona-free bovine oocytes were always located near the maternal chromosomes. Zona-free oocytes were enucleated by removing PB1 and about 3% of the adjacent oocyte cytoplasm without chromatin staining. Successful enucleation of zona-free bovine oocytes was achieved in 96.9% of cases. The rate of development to the blastocyst stage was significantly greater in embryos reconstructed from rapidly maturing oocytes (47.8%) than with oocytes maturing at 18.0-20.0 hpm (33.3%). Overall, two large groups of bovine oocytes could be distinguished during in vitro maturation by the time required to reach the second stage of metaphase. Bovine embryos reconstructed from rapidly maturing enucleated oocytes had a significantly greater rate of development to the blastocyst stage than did embryos derived from later-maturing oocytes. We conclude that blind enucleation is a simple and efficient method for preparing cytoplasts in zona-free bovine cloning.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 2","pages":"287-92"},"PeriodicalIF":0.0,"publicationDate":"2009-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2007.0090","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28230077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 8

Bioluminescence imaging of human embryonic stem cells transplanted in vivo in murine and chick models. 人胚胎干细胞在小鼠和鸡体内移植的生物发光成像。

Cloning Stem Cells Pub Date : 2009-06-01 DOI: 10.1089/clo.2008.0056

Helen Priddle, Anna Grabowska, Teresa Morris, Philip A Clarke, Andrew J McKenzie, Virginie Sottile, Chris Denning, Lorraine Young, Sue Watson

{"title":"Bioluminescence imaging of human embryonic stem cells transplanted in vivo in murine and chick models.","authors":"Helen Priddle, Anna Grabowska, Teresa Morris, Philip A Clarke, Andrew J McKenzie, Virginie Sottile, Chris Denning, Lorraine Young, Sue Watson","doi":"10.1089/clo.2008.0056","DOIUrl":"https://doi.org/10.1089/clo.2008.0056","url":null,"abstract":"Research into the behavior, efficacy, and biosafety of stem cells with a view to clinical transplantation requires the development of noninvasive methods for in vivo imaging of cells transplanted into animal models. This is particularly relevant for human embryonic stem cells (hESCs), because transplantation of undifferentiated hESCs leads to tumor formation. The present study aimed to monitor hESCs in real time when injected in vivo. hESCs were stably transfected to express luciferase, and luciferase expression was clearly detected in the undifferentiated and differentiated state. When transfected hESCs were injected into chick embryos, bioluminescence could be detected both ex and in ovo. In the SCID mouse model, undifferentiated hESCs were detectable after injection either into the muscle layer of the peritoneum or the kidney capsule. Tumors became detectable between days 10-30, with approximately a 3 log increase in the luminescence signal by day 75. The growth phase occurred earlier in the kidney capsule and then reached a plateau, whilst tumors in the peritoneal wall grew steadily throughout the period analysed. These results show the widespread utility of bioluminescent for in vivo imaging of hESCs in a variety of model systems for preclinical research into regenerative medicine and cancer biology.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 2","pages":"259-67"},"PeriodicalIF":0.0,"publicationDate":"2009-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0056","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28239438","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 15

Effect of PRL on in vitro follicle growth, in vitro oocyte maturation, fertilization and early embryonic development in mice. PRL对小鼠体外卵泡生长、体外卵母细胞成熟、受精及早期胚胎发育的影响。

Cloning Stem Cells Pub Date : 2009-06-01 DOI: 10.1089/clo.2008.0046

Erasmia Kiapekou, Dimitris Loutradis, George Mastorakos, Ritsa Bletsa, Panagiotis Beretsos, Evangelia Zapanti, Peter Drakakis, Aristeidis Antsaklis, Ann A Kiessling

{"title":"Effect of PRL on in vitro follicle growth, in vitro oocyte maturation, fertilization and early embryonic development in mice.","authors":"Erasmia Kiapekou, Dimitris Loutradis, George Mastorakos, Ritsa Bletsa, Panagiotis Beretsos, Evangelia Zapanti, Peter Drakakis, Aristeidis Antsaklis, Ann A Kiessling","doi":"10.1089/clo.2008.0046","DOIUrl":"https://doi.org/10.1089/clo.2008.0046","url":null,"abstract":"Prolactin (PRL), along with other hormones, plays a role in oocyte maturation, fertilization, and early embryonic development in mammals. In order to investigate the role of PRL on in vitro oocyte maturation from early follicular growth stages, as well as on fertilization and early embryonic development, we cultured preantral mouse follicles with and without PRL, followed by fertilization of the in vitro matured oocytes. Prolactin significantly improved the rate of oocyte maturation, fertilization, and early embryo development. Four isoforms of PRL-Receptor (R) have been found in whole ovaries of mice: one long (PRL-RL) and three short (-RS(1), -RS(2), and -RS(3)). We examined expression of the four PRL-R isoforms in preantral follicles, in cumulus-oocyte complexes (COCs) and in germinal vesicle GV stage oocytes by RT-PCR. Prolactin-RL, -RS(2) and -RS(3) mRNA, but not -RS(1), were expressed in preantral follicles, COCs, and GV stage oocytes. Our results indicate the prolactin pathway is functional in early preantral follicles, in COCs and in GV stage oocytes, and promotes oocyte maturation, meiosis, fertilization, and early embryonic development.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 2","pages":"293-300"},"PeriodicalIF":0.0,"publicationDate":"2009-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"28314994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 18

Demecolcine effects on microtubule kinetics and on chemically assisted enucleation of bovine oocytes. 去乙酰胆碱对牛卵母细胞微管动力学和化学辅助去核的影响。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0044

Naiara Zoccal Saraiva, Felipe Perecin, Simone Cristina Méo, Christina Ramires Ferreira, Tatiane Almeida Drummond Tetzner, Joaquim Mansano Garcia

{"title":"Demecolcine effects on microtubule kinetics and on chemically assisted enucleation of bovine oocytes.","authors":"Naiara Zoccal Saraiva, Felipe Perecin, Simone Cristina Méo, Christina Ramires Ferreira, Tatiane Almeida Drummond Tetzner, Joaquim Mansano Garcia","doi":"10.1089/clo.2008.0044","DOIUrl":"https://doi.org/10.1089/clo.2008.0044","url":null,"abstract":"This study aimed to evaluate the effect of demecolcine, a microtubule-depolymerizing agent, on microtubule kinetics; to determine the best concentration of demecolcine as a chemically assisted enucleation agent in metaphase I (MI) and metaphase II (MII) bovine oocytes, and to evaluate the embryonic development after nuclear transfer (NT) using chemically assisted enucleation of recipient oocytes. Oocytes in vitro matured for 12 h (MI) and 21 h (MII) were exposed to several concentrations of demecolcine and evaluated for enucleation or membrane protrusion formation. Demecolcine concentration of 0.05 microg/mL produced the highest rates of enucleation in group MI (15.2%) and protrusion formation in group MII (55.1%), and was employed in the following experiments. Demecolcine effect was seen as early as 0.5 h after treatment, with a significant increase in the frequency of oocytes with complete microtubule depletion in MI (58.9%) and MII (21.8%) compared to initial averages at 0 h (27.4% and 1.9%, respectively). Microtubule repolymerization was observed when MII-treated oocytes were cultured in demecolcine-free medium for 6 h (42.4% oocytes with two evident sets of microtubules). Chemically assisted enucleated oocytes were used as recipient cytoplasts in NT procedures to assess embryonic development. For NT, 219 of 515 oocytes (42.5%) formed protrusions and were enucleated, and reconstructed, resulting in 58 nuclear-transferred one-cell embryos. Cleavage (84.5%) and blastocyst development (27.6%) rates were assessed. In conclusion, demecolcine can be used at lower concentrations than routinely employed, and the chemically assisted enucleation technique was proven to be highly efficient allowing embryonic development in bovine.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"141-52"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0044","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27992697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 17

Beneficial effect of young oocytes for rabbit somatic cell nuclear transfer. 幼龄卵母细胞对兔体细胞核移植的有益作用。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0042

Fuliang Du, Jie Xu, Jifeng Zhang, Shaorong Gao, Mark G Carter, Chingli He, Li-Ying Sung, Sanjeev Chaubal, Rafael A Fissore, X Cindy Tian, Xiangzhong Yang, Y Eugene Chen

{"title":"Beneficial effect of young oocytes for rabbit somatic cell nuclear transfer.","authors":"Fuliang Du, Jie Xu, Jifeng Zhang, Shaorong Gao, Mark G Carter, Chingli He, Li-Ying Sung, Sanjeev Chaubal, Rafael A Fissore, X Cindy Tian, Xiangzhong Yang, Y Eugene Chen","doi":"10.1089/clo.2008.0042","DOIUrl":"https://doi.org/10.1089/clo.2008.0042","url":null,"abstract":"This study was designed to examine the effect of the age of rabbit oocytes on the developmental potential of cloned embryos. The metaphase II oocytes used for nuclear transfer (NT) were collected at 10, 12, 14, and 16 h post-hCG injection (hpi). The total number of oocytes collected per donor (21.4-23.7) at 12 to 16 hpi was similar, but significantly higher than that collected at 10 hpi (16.2). Additionally, a significant improvement in blastocyst development was achieved with embryos generated by electrically mediated cell fusion (56.0%), compared to those from nuclear injection (13.1 %) (Experiment 1). Markedly higher blastocyst development (45.8-54.5%) was also achieved with oocytes collected at 10-12 hpi than from those collected 14-16 hpi (8.3-14.3%) (Experiment 2). In Experiment 3, the blastocyst rates of NT embryos derived from oocytes harvested 12 hpi (39.2-42.8 %) were significantly higher than from those collected at 16 hpi (6.8-8.4 %) (p < 0.05), regardless of the donor cell age. Kinase activity assays showed variable changes of activity in rabbit oocytes over the period of 10-16 hpi; however, there was no correlation with preimplantational development (blastocyst rate vs. MPF, R = 0.326; blastocyst rate vs. MAPK, R = -0.131). Embryo transfer of NT embryos utilizing 12 hpi oocytes resulted in one full-term but stillborn, and one live cloned rabbit; thus, an efficiency of 1.7 % (n = 117) (Experiment 4). These results demonstrated that NT utilizing relatively young rabbit oocytes, harvested at 10-12 h after hCG injection, was beneficial for the development of NT embryos.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"131-40"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0042","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27905869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 21

Bovine ooplasm partially remodels primate somatic nuclei following somatic cell nuclear transfer. 体细胞核移植后，牛卵浆部分重塑灵长类体细胞核。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0061

Kai Wang, Zeki Beyhan, Ramon M Rodriguez, Pablo J Ross, Amy E Iager, German G Kaiser, Ying Chen, Jose B Cibelli

{"title":"Bovine ooplasm partially remodels primate somatic nuclei following somatic cell nuclear transfer.","authors":"Kai Wang, Zeki Beyhan, Ramon M Rodriguez, Pablo J Ross, Amy E Iager, German G Kaiser, Ying Chen, Jose B Cibelli","doi":"10.1089/clo.2008.0061","DOIUrl":"https://doi.org/10.1089/clo.2008.0061","url":null,"abstract":"Interspecies somatic cell nuclear transfer (iSCNT) has the potential to become a useful tool to address basic questions about the nucleus-cytoplasm interactions between species. It has also been proposed as an alternative for the preservation of endangered species and to derive autologous embryonic stem cells. Using chimpanzee/ bovine iSCNT as our experimental model we studied the early epigenetic events that take place soon after cell fusion until embryonic genome activation (EGA). Our analysis suggested partial EGA in iSCNT embryos at the eight-cell stage, as indicated by Br-UTP incorporation and expression of chimpanzee embryonic genes. Oct4, Stella, Crabp1, CCNE2, CXCL6, PTGER4, H2AFZ, c-MYC, KLF4, and GAPDH transcripts were expressed, while Nanog, Glut1, DSC2, USF2, Adrbk1, and Lin28 failed to be activated. Although development of iSCNT embryos did not progress beyond the 8- to 16-cell stage, chromatin remodeling events, monitored by H3K27 methylation, H4K5 acetylation, and global DNA methylation, were similar in both intra- and interspecies SCNT embryos. However, bisulfite sequencing indicated incomplete demethylation of Oct4 and Nanog promoters in eight-cell iSCNT embryos. ATP production levels were significantly higher in bovine SCNT embryos than in iSCNT embryos, TUNEL assays did not reveal any difference in the apoptotic status of the nuclei from both types of embryos. Collectively, our results suggest that bovine ooplasm can partially remodel chimpanzee somatic nuclei, and provides insight into some of the current barriers iSCNT must overcome if further embryonic development is to be expected.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"187-202"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0061","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27967971","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 41

Live birth of somatic cell-cloned rabbits following trichostatin A treatment and cotransfer of parthenogenetic embryos. 曲古斯汀A处理和孤雌胚胎共移植后体细胞克隆兔的活产。

Cloning Stem Cells Pub Date : 2009-03-01 DOI: 10.1089/clo.2008.0072

Qinggang Meng, Zsuzsanna Polgar, Jun Liu, Andras Dinnyes

{"title":"Live birth of somatic cell-cloned rabbits following trichostatin A treatment and cotransfer of parthenogenetic embryos.","authors":"Qinggang Meng, Zsuzsanna Polgar, Jun Liu, Andras Dinnyes","doi":"10.1089/clo.2008.0072","DOIUrl":"https://doi.org/10.1089/clo.2008.0072","url":null,"abstract":"Somatic cell nuclear transfer (SCNT) efficiency is still low in rabbit. Previous studies indicated that trichostatin A (TSA) treatment could improve cloning efficiency and term development in the mouse, and cotransfer of parthenogenetic (PA) embryos benefited the pregnancy of cloned embryos in porcine and the mouse. In this study we investigated the effect of TSA treatment on the term development of the SCNT rabbit embryos, and the possibility of the pregnancy maintenance of clones by cotransfer of PA embryos. The SCNT embryos were produced by fusing cumulus cells with enucleated cytoplasts before activation by electrical stimulation, and Dimethylaminopurine (6-DMAP) and Cyclohexamide (CHX) treatments. They were cultured in EBSS-complete medium regardless of their treatment with or without TSA. In vitro developmental data showed no differences in the cleavage and the blastocyst rates, and the blastocyst cell number between the TSA-treated and the untreated SCNT embryos. Two of the six recipients became pregnant after the embryo transfer (ET) in the TSA-treated group, and one pregnant female delivered seven live and three stillborn pups. The death of all live pups occurred within an hour to 19 days. Four of the seven recipients became pregnant in the TSA-untreated group. Three of them gave birth to six live and eight stillborn pups. Four pups of the TSA-untreated group have grown into adulthood, and three of them produced progeny. Cotransfer of three to four PA embryos with 26-32 SCNT embryos to the same recipient resulted in pregnancy and birth rates statistically no different compared to the control SCNT ET group. In conclusion, our results indicate that TSA treatment has a limited effect on the in vitro development of the SCNT embryos; furthermore, both the TSA-treated and the untreated clones can develop to term in rabbits, but none of the offspring from TSA-treated embryos survived to adulthood in our experiment.","PeriodicalId":49217,"journal":{"name":"Cloning Stem Cells","volume":"11 1","pages":"203-208"},"PeriodicalIF":0.0,"publicationDate":"2009-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/clo.2008.0072","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"27968432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 92