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Production of Cloned Mice by Nuclear Transfer of Cumulus Cells

BACKGROUND: Over the past several years, mammals have been successfully cloned by either the splitting of an early stage embryo or nuclear transfer of adult somatic cells (NT) into oocytes. Although it has been 15 years since the generation of the first cloned mammals from somatic cells by NT, the s...

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Autores principales: Kurd, Soleiman, Zarei, Mohammad Ali, Fathi, Fardin, Ghadimi, Tayyeb, Hakhamaneshi, Mohammad Saeed, Jalili, Ali
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Avicenna Research Institute 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3732868/
https://www.ncbi.nlm.nih.gov/pubmed/23919122
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author Kurd, Soleiman
Zarei, Mohammad Ali
Fathi, Fardin
Ghadimi, Tayyeb
Hakhamaneshi, Mohammad Saeed
Jalili, Ali
author_facet Kurd, Soleiman
Zarei, Mohammad Ali
Fathi, Fardin
Ghadimi, Tayyeb
Hakhamaneshi, Mohammad Saeed
Jalili, Ali
author_sort Kurd, Soleiman
collection PubMed
description BACKGROUND: Over the past several years, mammals have been successfully cloned by either the splitting of an early stage embryo or nuclear transfer of adult somatic cells (NT) into oocytes. Although it has been 15 years since the generation of the first cloned mammals from somatic cells by NT, the success rate for producing live offspring by this technique is low regardless of the cell type and animal species used. However, these techniques have the potential to be important tools for future research in basic biology. In the present study, we described our experiences in producing successfully cloned mouse using NT method and piezo-actuated micromanipulator. METHODS: B6D2F1 mice, 8-12 weeks old, were superovulated with injections of 5 IU of pregnant mare serum gonadotropin and 5 IU of human chorionic gonadotropin administered 48 hr apart. Enucleation and donor nuclei cumulus cell injection were performed with a piezo-actuated micromanipulator after which activation and trichostatin A treatment were used for reconstructed oocytes. Two-cell stage cloned embryos that developed in the mWM medium were transferred into the oviducts of pseudopregnant NMRI mice. RESULTS: Of 367 oocytes collected, 131 (69%) developed into 2-cell stage embryos. Of these, 5 (1%) live pups were successfully delivered. We used NMRI foster mother to raise the pups by lactation. One adult cloned mouse was mated, after which she delivered and raised normal offspring. CONCLUSION: For mouse cloning, the present study also successfully tested the capability of somatic cell nuclear transfer SCNT using a piezo unit.
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spelling pubmed-37328682013-08-05 Production of Cloned Mice by Nuclear Transfer of Cumulus Cells Kurd, Soleiman Zarei, Mohammad Ali Fathi, Fardin Ghadimi, Tayyeb Hakhamaneshi, Mohammad Saeed Jalili, Ali Avicenna J Med Biotechnol Original Article BACKGROUND: Over the past several years, mammals have been successfully cloned by either the splitting of an early stage embryo or nuclear transfer of adult somatic cells (NT) into oocytes. Although it has been 15 years since the generation of the first cloned mammals from somatic cells by NT, the success rate for producing live offspring by this technique is low regardless of the cell type and animal species used. However, these techniques have the potential to be important tools for future research in basic biology. In the present study, we described our experiences in producing successfully cloned mouse using NT method and piezo-actuated micromanipulator. METHODS: B6D2F1 mice, 8-12 weeks old, were superovulated with injections of 5 IU of pregnant mare serum gonadotropin and 5 IU of human chorionic gonadotropin administered 48 hr apart. Enucleation and donor nuclei cumulus cell injection were performed with a piezo-actuated micromanipulator after which activation and trichostatin A treatment were used for reconstructed oocytes. Two-cell stage cloned embryos that developed in the mWM medium were transferred into the oviducts of pseudopregnant NMRI mice. RESULTS: Of 367 oocytes collected, 131 (69%) developed into 2-cell stage embryos. Of these, 5 (1%) live pups were successfully delivered. We used NMRI foster mother to raise the pups by lactation. One adult cloned mouse was mated, after which she delivered and raised normal offspring. CONCLUSION: For mouse cloning, the present study also successfully tested the capability of somatic cell nuclear transfer SCNT using a piezo unit. Avicenna Research Institute 2013 /pmc/articles/PMC3732868/ /pubmed/23919122 Text en Copyright © 2013 Avicenna Research Institute http://creativecommons.org/licenses/by-nc/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.
spellingShingle Original Article
Kurd, Soleiman
Zarei, Mohammad Ali
Fathi, Fardin
Ghadimi, Tayyeb
Hakhamaneshi, Mohammad Saeed
Jalili, Ali
Production of Cloned Mice by Nuclear Transfer of Cumulus Cells
title Production of Cloned Mice by Nuclear Transfer of Cumulus Cells
title_full Production of Cloned Mice by Nuclear Transfer of Cumulus Cells
title_fullStr Production of Cloned Mice by Nuclear Transfer of Cumulus Cells
title_full_unstemmed Production of Cloned Mice by Nuclear Transfer of Cumulus Cells
title_short Production of Cloned Mice by Nuclear Transfer of Cumulus Cells
title_sort production of cloned mice by nuclear transfer of cumulus cells
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3732868/
https://www.ncbi.nlm.nih.gov/pubmed/23919122
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