2011 (SBTE) 25th Annual Meeting Proceedings - International ...

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D.P .P.A.F .A.F. Braga & E. Bor orges Jr. 2011. The Role of Veterinarians in Human In Vitro Embryo Production. SSSSSSSSSSS ssssssssssssssssssssssss Acta Scientiae Veterinariae. 39(Suppl 1): s73 - s80. 58 Wang W.H., Meng L., Hackett R.J., Odenbourg R. & Keefe D.L. 2001. The spindle observation and its relationship with fertilization after intracytoplasmic sperm injection in living human oocytes. Fertility and Sterility. 75(2): 348-353. 59 Wang W.H., Meng L., Hackett R.J. & Keefe D.L. 2001. Developmental ability of human oocytes with or without birefringent spindles imaged by Polscope before insemination. Human Reproduction. 16(7): 1464-1468. 60 Wilding M., Di Matteo L., D’Andretti S., Montanaro N., Capobianco C. & Dale B. 2007. An oocyte score for use in assisted reproduction. Journal of Assisted Reproduction and Genetics. 24(8): 350-358. 61 Wright J.M. 1981. Non-surgical embryo transfer in cattle embryo-recipient interactions. Theriogenology. 15(1): 43-56. www.ufrgs.br/actavet 39(Suppl 1) s80
M.E.F .F. Oliv liveir eira. 2011. Estado da arte da superovulação em ovelhas. aaaaaa aaaaaaaaaaaa aaaaaaaaaaaa Acta Scientiae Veterinariae. 39(Supl 1): s1 - s7. Acta Scientiae Veterinariae, 2011. 39(Suppl1): s81. ISSN 1679-9216 (Online) Vet eter erinar inary applications in regener egenerativ tive medicine; developmen elopment t of induced pluripotential stem cells (iPSC) in dogs Sehw ehwon Koh, Steve e Bischoff ischoff, Shengdar Tsai & Jor orge A. Piedr iedrahita ABSTRACT Background: Pluripotent stem cells such as embryonic stem cells can give rise to derivatives of all three germ layers and thus have great potential for clinical applications related to regenerative medicine. By analyzing totipotent or pluripotent gene expression signatures from eggs, embryonic blastomeres and embryonic stem cells, Takahashi & Yamanaka (2006) developed a method to directly rewire the circuitry of adult somatic cells to pluripotency by transfection with a range of transcription factors. Ectopic expression of four transcription factors Oct4, Klf4, Sox2 and c-Myc (OKSM) was capable of resetting the adult somatic cell to pluripotency. This approach has now been applied to a range of mammalian species including mice, humans, horses and pigs. Our interest is the application of this technology to the dog both for clinical application in veterinary medicine and as a way to understanding some of the limitations and strengths of this technology when applied to humans. Review: Here we report the derivation of iPS cells from adult canine fibroblast by retroviral OSKM transduction. The isolated canine iPS cells were expanded in three different iPS culture media (FGF2, LIF and FGF2 plus LIF) and only the cells cultured in FGF2 plus LIF showed strong AP activity and to express pluripotency markers, POU5F1 (OCT4), SOX2, NANOG and LIN28 as well as ES cells-specific genes (PODXL, DPPA5, FGF5, REX1 and LAMP1). To determine the ability of the cell to differentiate into derivatives of all tree germ layers; endoderm, mesoderm and ectoderm, we utilized both embryoid body formation and directed differentiation using chemical inducers. In vitro differentiation by formation of embryoid bodies (EBs) and directed differentiation showed cell derivatives of all three germ layers as confirmed by expression for AFP, CXCR4 and SOX17 N (endoderm), desmin (DES), vimentin (VIM), MSX1 and BMP2 (mesoderm) and glial fibrillary acidic proten (GFAP), TUJ1, NCAM and bIII-tubulin (TUBB, ectoderm). In vivo differentiation was tested by development of teratomas after injection into immunodeficient mice. Results indicated that the putative canine iPS cells were capable of creating solid tumors that expressed markers for all three germ layers. Conclusion: Embryonic stem cells have tremendous potential in the area of regenerative medicine but the difficulties in their isolation, and the inability to rapidly and efficiently develop lines that can be genetically matched to the recipient, reduces their clinical usefulness. In contrast, pluripotential stem cells (iPS) developed through direct reprogramming with transcription factors allow the derivation of patient-derived stem cells. This will allow the development of cell lines that can be used in the area of regenerative medicine in both human and veterinary medicine. However, there are still some issues of stability and culture requirements that must be elucidated before this technology can be fully applied in the clinics. Keywords: induced pluripotent stem cells, stem cells, dogs, regenerative medicine. Center for Comparative Medicine and Translational Research and Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University. CORRESPONDENCE: J.A. Piedrahita [jorge_piedrahita@ncsu.edu]. Center for Comparative Medicine and Translational Research and Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 4700 Hillsborough St., ZIP CODE: 27606, Raleigh, NC, U.S.A. s81
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A262 Bovine Oocyte Vitrification: E
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