Biotechnology1682-296x1682-2978Asian Network for Scientific Information10.3923/biotech.2007.373.382in vitro Matured Oocytes of Heterogeneous Quality Through Intracytoplasmic Sperm Injection Technique
]]>RahmanAbu Nasar Md. Aminoor AbdullahRamli Bin EmbongWan Khadijah Wan 3200763Experiments were conducted to develop an intracytoplasmic
sperm injection (ICSI) protocol to produce goat embryos from in vitro
matured heterogeneous oocytes. Two different protocols (‘Protocol
1 or P-1 and ‘Protocol 2
or P-2) were formulated and tested to develop a suitable one. Cumulus-oocyte
complexes (COCs) were obtained either by LOPU (P-1) or by ovariectomy and slicing
(P-2). Recovered COCs were cultured in a microdrop (50 μL) of HEPES-buffered
in vitro maturation (IVM) medium with 10% oestrus goat serum (OGS) (P-1)
or IVM medium without HEPES with 15% OGS (P-2) in presence of CO2 (5%)
at 38.5°C either for 24 h (P-1) or 27 h (P-2). After IVM, COCs were denuded
to select a mature oocyte (MII) and held in the incubator in presence of CO2
(5%) until ICSI. Motile sperm were selected from frozen-thawed buck semen by
swim-up either in mDM (P-1) or mSOF (P-2) medium. A motile sperm was immobilized
and injected, head-first, into the ooplasm of a MII oocyte. Sham injections
were also performed as control. Injected oocytes were cultured in mSOF medium
either with 10% (P-1) or 20% (P-2) goat serum in presence of CO2 (5%)
at 38.5°C. A significantly higher (p<0.01) number of COCs was recovered
with P-2 (17.5 COCs/donor) than P-1 (6.7 COCs/donor). However, the quality of
the COCs was better with P-1 than P-2. A higher proportion of COCs were matured
with P-2 (69.3%) than P-1 (55.5%), which was not statistically significant.
A fertilization rate of 28.6% was obtained in P-1 without any cleavage. Fertilization
rate (54.1%) was significantly higher (p<0.05) in P-2 with a cleavage rate
of 18.2% and few morula-stage embryos were obtained (2.9%). In conclusion, better
fertilization and embryo development rates were obtained with P-2 than P-1.
Morula-stage embryos were obtained using ICSI technique from in vitro
matured heterogeneous goat oocytes without any artificial activation and first
reported in Malaysia.]]>Amir, A.A.B.,2007in vitro maturation, fertilization and culture (IVMFC) techniques.]]>2007Baldassarre, H., D.G. de Matos, C.C. Furnus, T.E. Castro and E.I. Cabrera Fische,199435145150Baldassarre, H., B. Wang, N. Kafidi, C. Keefer, A. Lazaris and C.N. Karatzas2002in vitro embryo production technologies.]]>57275284Baldassarre, H., C. Keefer, B. Wang, A. Lazaris and C.N. Karatzas, 2003in vitro matured oocytes recovered by laparoscopic ovum pick-up. ]]>5279285Baldassarre, H. and C.N. Karatzas,200482-83255266Brackett, B.G. and G. Oliphant,1975in vitro.]]>12260274Hamano, K., X. Li, X.Q. Qian, K. Funauchi, M. Furudate and Y. Minat, 19996011941197Hamberger, L., A. Sjogren, K. Lundin, B. Soderlund and L. Nilsson et al.,19957263268Jamaluddin, A. and V.I.H. Ng,20051057Jimenez-Macedo, A.R., D. Izquierdo, B. Anguita and M.T. Paramio,2005in vitro fertilization employing oocytes derived from prepubertal goats.]]>6412491262Jimenez-Macedo, A.R., B. Anguita, D. Izquierdo, T. Mogas and M.T. Paramio,20066610651072Kasai, T., K. Hoshi and R. Yanagimachi,19997187193Keskintepe, L., G.C. Luvoni, S.J. Rzucidlo and B.G. Brackett,1996in vitro production of viable uterine stage caprine embryos.]]>20247254Keskintepe, L., P. Morton, S.E. Smith, M.J. Tucker, A.A. Simplicio and B.G. Brackett,19975261265Nicholas, F.W., 199642205214Ongeri, E.M., C.L. Bormann, R.E. Butler, D. Melican and W.G. Gavin et al.,2001in vitro fertilization and parthenogenetic activation by different methods.]]>5519331945Perry, A.C., T. Wakayama and R. Yanagimachi,199960747755Phua, A.C.Y.,2006in vitro maturation, fertilization and culture techniques.]]>2006Simplicio, A.A., B.G. Brackett and L. Keskintepe,1997in vitro fertilization (IVF).]]>47299299Takahashi, Y. and N.L. First,1992In vitro development of bovine one-cell embryos: Influence of glucose, lactate, pyruvate, amino acids and vitamins.]]>37963978Wang, B., H. Baldassarre, J. Pierson, F. Cote, K.M. Rao and C.N. Karatzas,2003in vitro and in vivo development of goat embryos produced by intracytoplasmic sperm injection using tail-cut spermatozoa.]]>11219227Zhou, J.B., Y.G. Wu, D. Han, L.Q. Liu and X.W. Tan et al.,200437367374, (In Chinese)