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Genetically corrected ES cells derived from cloned embryos of infertile mice - a potential application for therapeutic cloning

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Abstract:

The ability of embryonic stem (ES) cells to differentiate into germ cells in culture has been established, and functional germ cells, including haploid cells, have been generated. As with many genetic diseases, generation of the missing cell type in vitro for male infertility provides a valuable tool for studying the mechanism of the disease and discovering therapies. Toward this goal, our lab has been applying the somatic cell nuclear transfer (SCNT) technology to develop treatments for genetic disease using the mouse as a model. In the present study, we aimed to correct a mouse germ cell deficiency which renders male infertility. First, we generated cloned embryos using skin fibroblast cells of the infertile mouse, WBB6F1/J-KitW/KitW-v (W/W-v). We then derived nuclear transfer ES cells (ntES) from the cloned embryos. The ntES cell lines carrying this genetic deficiency were characterized for their pluripotency and their genetic deficiency was corrected by gene targeting. Lastly, we determined the potential of the genetically corrected ntES cells for their in vitro ability to differentiate into functional male germ cells and compare that to control ES cell lines. Experiments for in vivo assay of the functions of genetically corrected ntES cells by producing ntES cell-derived animals through tetraploid complementation or second round of SCNT are on-going.
Our research will ultimately lead to not only the production of offspring from infertile mice and advance the study of therapeutic cloning as an alternative assisted reproduction technology (ART) to germ cell therapy in humans, but also offers the unique approach to study cell reprogramming in stem cell - germ cell transformation.
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Association:
Name: Connecticut's Stem Cell Research International Symposium
URL:
http://stemconn.org


Citation:
URL: http://citation.allacademic.com/meta/p319304_index.html
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MLA Citation:

Amano, Tomokazu., Ma, Yinghong., He, Zuping., Amano, Misa., Treaster, Stephen., Lin, Chih-Jen., Dym, Martin., Tian, Cindy. and Yang, Jerry. "Genetically corrected ES cells derived from cloned embryos of infertile mice - a potential application for therapeutic cloning" Paper presented at the annual meeting of the Connecticut's Stem Cell Research International Symposium, Omni Hotel, New Haven, CT, Mar 23, 2009 <Not Available>. 2014-11-29 <http://citation.allacademic.com/meta/p319304_index.html>

APA Citation:

Amano, T. , Ma, Y. , He, Z. , Amano, M. , Treaster, S. , Lin, C. , Dym, M. , Tian, C. and Yang, J. , 2009-03-23 "Genetically corrected ES cells derived from cloned embryos of infertile mice - a potential application for therapeutic cloning" Paper presented at the annual meeting of the Connecticut's Stem Cell Research International Symposium, Omni Hotel, New Haven, CT <Not Available>. 2014-11-29 from http://citation.allacademic.com/meta/p319304_index.html

Publication Type: Poster
Review Method: Peer Reviewed
Abstract: The ability of embryonic stem (ES) cells to differentiate into germ cells in culture has been established, and functional germ cells, including haploid cells, have been generated. As with many genetic diseases, generation of the missing cell type in vitro for male infertility provides a valuable tool for studying the mechanism of the disease and discovering therapies. Toward this goal, our lab has been applying the somatic cell nuclear transfer (SCNT) technology to develop treatments for genetic disease using the mouse as a model. In the present study, we aimed to correct a mouse germ cell deficiency which renders male infertility. First, we generated cloned embryos using skin fibroblast cells of the infertile mouse, WBB6F1/J-KitW/KitW-v (W/W-v). We then derived nuclear transfer ES cells (ntES) from the cloned embryos. The ntES cell lines carrying this genetic deficiency were characterized for their pluripotency and their genetic deficiency was corrected by gene targeting. Lastly, we determined the potential of the genetically corrected ntES cells for their in vitro ability to differentiate into functional male germ cells and compare that to control ES cell lines. Experiments for in vivo assay of the functions of genetically corrected ntES cells by producing ntES cell-derived animals through tetraploid complementation or second round of SCNT are on-going.
Our research will ultimately lead to not only the production of offspring from infertile mice and advance the study of therapeutic cloning as an alternative assisted reproduction technology (ART) to germ cell therapy in humans, but also offers the unique approach to study cell reprogramming in stem cell - germ cell transformation.


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