scholarly journals Remodeling somatic nuclei via exogenous expression of protamine 1 to create spermatid-like structures for somatic nuclear transfer

2016 ◽  
Vol 11 (11) ◽  
pp. 2170-2188 ◽  
Author(s):  
Marta Czernik ◽  
Domenico Iuso ◽  
Paola Toschi ◽  
Saadi Khochbin ◽  
Pasqualino Loi
Keyword(s):  
Nuclear Transfer ◽  
Exogenous Expression ◽  
Somatic Nuclear Transfer ◽  
Protamine 1

Effect of telophase enucleation on bovine somatic nuclear transfer

2000 ◽  
Vol 54 (6) ◽  
pp. 989-998 ◽  
Author(s):  
J.-L. Liu ◽  
M.-K. Wang ◽  
Q.-Y. Sun ◽  
Z. Xu ◽  
D.-Y. Chen
Keyword(s):  
Nuclear Transfer ◽  
Somatic Nuclear Transfer

67 GENERATION OF PIGS TRANSGENIC FOR hCD59/DAF AND HUMAN THROMBOMODULIN BY SOMATIC NUCLEAR TRANSFER

2006 ◽  
Vol 18 (2) ◽  
pp. 142 ◽  
Author(s):  
B. Petersen ◽  
W. Kues ◽  
A. Lucas-Hahn ◽  
A.-L. Queisser ◽  
E. Lemme ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Fluorescein Isothiocyanate ◽  
Coagulation Cascade ◽  
Cell Cycle Synchronization ◽  
Novel Approach ◽  
Donor Cells ◽  
Serum Gonadotropin ◽  
Somatic Nuclear Transfer ◽  
Porcine Organs

After a porcine–to–primate xenotransplantation, hyperacute rejection (HAR) destroys the transplanted organ within minutes. The HAR can be overcome either by a knockout of the gene for α–1,3–galactosyltransferase or by overexpression of human complement regulatory proteins such as hCD59 and DAF. When HAR can be controlled, the next hurdle is acute vascular rejection (AVR) which is primarily due to an incompatibility of human protein C and porcine thrombomodulin, both of which are important factors in the coagulation cascade. This incompatibility leads to thrombosis and finally to a disseminated intravascular coagulation (DIC), causing rejection of the xenotransplant. Human thrombomodulin is a good candidate gene for improving survival time of porcine organs after xenotransplantation and for overcoming AVR. Here, we transfected adult fibroblasts obtained from a double transgenic boar (hCD59/DAF) with a construct for human thrombomodulin (hTM). After selection with 800 μg/mL G418 for 14 days, cells were analyzed for integration of the construct by PCR and were visually selected for expression of the hTM–GFP fusion protein under ultraviolet light with a fluorescein isothiocyanate (FITC) filterset. A total of 39 positve clones were obtained, of which two were used in somatic nuclear transfer. Ovaries were collected from a local slaughterhouse, and follicles of 2–5 mm in diameter were aspirated. After 38–42 h of in vitro maturation oocytes were denuded and enucleated. For cell cycle synchronization, the donor cells were serum–starved for 48 h, subsequently trypsinized and placed into the perivitelline space of the enucleated oocytes. The complexes were fused and activated electrically followed by an incubation in DMAP for 3 h. Puberal gilts were synchronized by treatment with 5 mL Regumate® (Intervet UK, Ltd., Milton Keynes, Buckinghamshire, UK) for 13 days. At the end of treatment, the animals received 1000 IU pregnant mare serum gonadotropin (PMSG) intramuscularly followed by an injection of 500 IU hCG 72 h later. Cloned embryos were transferred surgically 20 h after hCG injection. Maintenance of pregnancy was supported by injections of 1000 IU PMSG on Day 11 and 500 IU hCG on Day 14 of the pregnancy. Out of 1409 reconstructed complexes, 1161 were fused (82.4%) successfully. In total, 1040 embryos were transferred to 8 recipients (∼130 embryos/gilt, range 70–162). Five of the eight recipients (62.5%) became pregnant as determined by ultrasound on Days 25, 36, and 51 and will farrow within the next weeks. These results show that cloning with triple transgenic adult donor cells is compatible with high pregnancy rates. Porcine multitransgenic organs will be used in perfusion experiments to test the effectiveness of this novel approach to overcoming the incompatibilities between the porcine and the human coagulation systems. This project is funded by the Deutsche Forschungsgemeinschaft (FOR 535). The hTM–construct was a gift of Dr. Wu which is gratefully acknowledged.

scholarly journals Successful Mouse Cloning of an Outbred Strain by Trichostatin A Treatment after Somatic Nuclear Transfer

2007 ◽  
Vol 53 (1) ◽  
pp. 165-170 ◽  
Author(s):  
Satoshi KISHIGAMI ◽  
Hong-Thuy BUI ◽  
Sayaka WAKAYAMA ◽  
Kenzo TOKUNAGA ◽  
Nguyen Van THUAN ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Trichostatin A ◽  
Outbred Strain ◽  
Somatic Nuclear Transfer

Response to “May a Woman Clone Herself” by Jean Chambers (CQ Vol 10, No 2)

2002 ◽  
Vol 11 (1) ◽  
pp. 83-86
Author(s):  
Timothy F. Murphy
Keyword(s):  
Nuclear Transfer ◽  
Assisted Reproductive Technologies ◽  
Married Couples ◽  
Reproductive Technologies ◽  
Same Sex ◽  
Reproductive Cloning ◽  
Same Sex Couples ◽  
Human Reproductive ◽  
Somatic Nuclear Transfer ◽  
Do So

For many commentators in bioethics and the law, safety is the fulcrum for evaluating the ethics of human reproductive cloning. Carson Strong has argued that if cloning were effective and safe it should be available to married couples who have tried to have children through various assisted reproductive technologies (ARTs) but been unable to do so. On his view, cloning should be available only as reproductive last resort. I challenged that limited use by trying to show that the arguments Strong adduces in favor of reproductive somatic nuclear transfer (SNT) for married couples extend to same-sex couples as well, who face a different kind of infertility. I also went on to argue that his justifications would in fact extend the legitimate use of SNT to any couples regardless of whether they had fertility difficulties or not.

scholarly journals Somatic nuclear transfer in livestock species

2001 ◽  
Vol 44 (4) ◽  
pp. 351-364 ◽  
Author(s):  
B. Kühholzer ◽  
G. Brem
Keyword(s):  
Somatic Cell ◽  
Somatic Cell Nuclear Transfer ◽  
Nuclear Transfer ◽  
Transgenic Animals ◽  
Cell Types ◽  
Powerful Technique ◽  
Livestock Species ◽  
Recent Advances ◽  
Somatic Nuclear Transfer ◽  
Reported Data

Abstract. In this review, we discuss the recent advances in somatic cell nuclear transfer (NT) in sheep, cattle, goats, swine and rabbits. Numerous advances have been reported as this technique has developed over the last five years. In the first part of this review, we describe the reported data pertaining to each of the species mentioned above. Theories are offered to explain the different results seen between different species and cell types. One of the main aspects of somatic cell NT, the production of transgenic animals will also be reviewed. Future applications of this powerful technique are discussed. This review concludes with a discussion of some of the problems observed in animals produced using NT as well as possible Solutions for these challenges.

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