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non-human primates after transplantation with hCD46 transgenic porcine islets. American Journal of Transplantation, 9(12), 2716-2726. Vatzias, G. and Hagen, D. R. (1999) Effects of porcine follicular fluid and oviduct-conditioned media on maturation and fertilization of porcine oocytes in vitro. Biology of Reproduction, 60(1), 42-48. Velander, W. H., Johnson, J. L., Page, R. L., Russell, C. G, Subramanian, A., Wilkins, T. D., Gwazdauskas, F. C., Pittius, C. and Drohan, W. N. (1992) High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C. Proceedings of the National Academy of Sciences of the USA, 89(24), 12003-12007. Wakayama, S., Ohta, H., Hikichi, T., Mizutani, E., Iwaki, T., Kanagawa, O. and Wakayama, T. (2008) Production of healthy cloned mice from bodies frozen at -20 degrees C for 16 years. Proceedings of the National Academy of Sciences of the USA, 105(45), 17318-17322. Wakayama, T., Perry, A. C., Zuccotti, M., Johnson, K. R. and Yanagimachi, R. (1998) Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature, 394(6691), 369-374. Wakayama, T., Rodriguez, I., Perry, A. C., Yanagimachi, R., Mombaerts, P. (1999) Mice cloned from embryonic stem cells. Proceedings of the National Academy of Sciences of the USA, 96(26), 14984-14989. Walker, S. C., Shin, T., Zaunbrecher, G. M., Romano, J. E., Johnson, G. A., Bazer, F. W. and Piedrahita, J. A. (2002) A highly efficient method for porcine cloning by nuclear transfer using in vitro-matured oocytes. Cloning and Stem Cells, 4(2), 105-112. Walters, E. M. and Wheeler, M. B. (2002) Parthenogenetic activation of in vitro-matured pig oocytes with the combination of calcium ionophore (A23 187) and cycloheximide. Theriogenology, 57(1), 709. Wang, W. H., Abeydeera, L. R., Cantley, T. C. and Day, B. N. (1997a) Effects of oocyte maturation media on development of pig embryos produced by in vitro fertilization. Journal of Reproduction and Fertility, 111(1), 101-108. Wang, W. H., Sun, Q. Y., Hosoe, M. and Shioya, Y. (1997b) Calcium- and meiotic spindle-independent activation of pig oocytes by the inhibition of staurosporine-sensitive protein kinase. Zygote, 5(1), 75-82. Wani, N. A., Wernery, U., Hassan, F. A., Wernery, R. and Skidmore, J. A. (2010) 148
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The University of -210 Nottingham M
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treated oocytes was recorded at 38
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TABLE OF CONTENTS ABSTRACT ........
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2.5.1.2 Preparation of enucleation/
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CHAPTER 6 .........................
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LIST OF FIGURES Figure 1.1 Process
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LIST OF ABBREVIATIONS A23187 AC AI
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MgSO4.7H20 MAPK MAPKK MAPKKK MBP mg
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CHAPTER 1 LITERATURE REVIEW The aim
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The production of multiple offsprin
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cytoplasm which contained the nucle
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pluripotency, as demonstrated by th
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gene-targeting via somatic cell clo
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conventional breeding and genetic m
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Oocyte maturation is generally cont
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ooýcýýommý MPF Time Figure 1.3
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not obtain clones by SCNT using bra
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A Nuclear NEBD PCC reformation DNA
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1.2.3.2 MPF, MAPK, NEBD and PCC in
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Little is known about the mechanism
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development of porcine embryos prod
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short-wavelength, UV excitable fluo
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The procedure is to expel the PBI a
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The majority of successful porcine
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Until 1997, the established dogma i
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deacetylases (HDACs). Trichostatin
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Figure 1.5 presents the general exp
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CHAPTER 2 General materials and met
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2.3 Oocyte staining and assessment
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holding pipette enucleation pipette
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! 00-0 !".. ww "'') ^ýýe ;ý ý
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,rjý. 7!: rY Is All Pi D r. Figure
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were washed three times in embryo c
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Three replicates of batches of oocy
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fertilisation, the occurrence of po
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3.2 MATERIALS AND METHODS 3.2.1 Exp
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I 57
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3.3 RESULTS 3.3.1 Conventional matu
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ýIC M 00 N N N N 0 öa oÜd U 0 0
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Meiotic stage in CAMP treated oocyt
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3.4 DISCUSSION The development of N
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44 h. It was comparable to that (93
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oocyte (Chen et al., 1990). In the
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CHAPTER 4 Parthenogenetic developme
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mechanical and chemical procedures.
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COCs were collected and oocytes wer
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f. " ý1403: 0.71 14 - 'Ir I A Figu
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100% 90% 80% 70% d 60% 50% 0 40% 30
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4.4 DISCUSSION For successful porci
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In addition to the listed benefits
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CHAPTER 5 Effect of caffeine treatm
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in vitro substrates. In pigs, Naito
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5.2 MATERIALS AND METHODS 5.2.1 Exp
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5.2.2 In vitro maturation of porcin
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Blastocysts at day 7 were stained a
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Figure 5.2 MPF and MAPK activities
Page 113 and 114: A 4500000 N 4000000 3500000 Z 30000
Page 115 and 116: e ýo; +ý \.. 4.0 ob "i "s NW Ir
Page 117 and 118: 5.3.4 Effects of caffeine treatment
Page 119 and 120: Figure 5.6 Effects of 5 mM caffeine
Page 121 and 122: 5.4 DISCUSSION In the first experim
Page 123 and 124: In the final experiment, developmen
Page 125 and 126: CHAPTER 6 General discussion 7.1 Ma
Page 127 and 128: cumulus cells could be observed spe
Page 129 and 130: parthenogenetic development excludi
Page 131 and 132: demonstrated treatment of the TI en
Page 133 and 134: REFERENCES Abeydeera, L. R., Wang,
Page 135 and 136: Bromhall, J. D. (1975) Nuclear trna
Page 137 and 138: Christmann, L., Jung, T. and Moor,
Page 139 and 140: and human ovarian oocytes. Nature,
Page 141 and 142: Funahashi, H., Cantley, T. C. and D
Page 143 and 144: Hartwell, L. H. (1973) Three additi
Page 145 and 146: Jelinkovä, L., Kubelka, M., Motlik
Page 147 and 148: Kubelka, M., Anger, M., Pavlok, A.,
Page 149 and 150: Le Bourhis, D., Beaujean, N., Ruffi
Page 151 and 152: Liu, L., Oldenbourg, R., Trimarchi,
Page 153 and 154: Minshull, J., Blow, J. J. and Hunt,
Page 155 and 156: Oback, B. and Wells, D. N. Cloning
Page 157 and 158: A. E., Garst, A. S., Moore, M., Dem
Page 159 and 160: Rossomando, A. J., Payne, D. M., We
Page 161 and 162: Spemann, H. Embryonic development a
Page 163: Development. 55(2), 121-127. Törne
Page 167 and 168: oocytes. Reproduction, 125(5), 645-
Page 169 and 170: 00 Cl) h 'Cb a) Ü^W wv F NC a b d
Page 171 and 172: In In ý. ö O V %) ö ö N _ ö O
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Page 175 and 176: aý yy U a 2 w I-N O N 'T N 1-1 NO
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