Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/131312
Title: Intracytoplasmic sperm injection of mouse oocytes with 5 mM Ca2+ at different intervals
Authors: Ahmadi, A.A.
Ng, S.C. 
Liow, S.L. 
Ali, J. 
Bongso, A. 
Ratnam, S.S. 
Keywords: Ca2+ injection
Intracytoplasmic sperm injection
Micro-injection
Mouse oocyte
Spermatozoa
Issue Date: 1995
Source: Ahmadi, A.A., Ng, S.C., Liow, S.L., Ali, J., Bongso, A., Ratnam, S.S. (1995). Intracytoplasmic sperm injection of mouse oocytes with 5 mM Ca2+ at different intervals. Human Reproduction 10 (2) : 431-435. ScholarBank@NUS Repository.
Abstract: The objective of this investigation was to determine whether intracytoplasmic sperm injection (ICSI) can be performed in the mouse. Metaphase II oocytes were obtained from F1 hybrid mice (C57BL x CBA) by i.p. injections of 10 IU pregnant mare's serum gonadotrophin (PMSG) and human chorionic gonadotrophin (HCG) administered 48 h apart. Oocytes with cumulus oophorus were retrieved 13-14 h post HCG. Cumulus was dispersed with 0.1% hyaluronidase. Mouse spermatozoa were obtained from the cauda epididymides of males of the same strain. The spermatozoa were processed by the standard swim-up procedure. The harvested spermatozoa were then incubated for 1.5 h to allow capacitation. Healthy oocytes were injected with 3-4 pl 5 mM Ca2+, followed by one live morphologically normal spermatozoon into the cytoplasm at intervals of 0, 0.5, 1, 2 and 3 h. The proportion of 2-cell embryos that developed from oocytes injected with Ca2+ and spermatozoa ranged between 29.5 and 36.5% in all groups, with no statistical difference between treatments. Chromosomal analysis showed that two-thirds of the ICSI-derived 2-cell embryos were diploid. The proportion of pathenogenetically activated embryos in the ICSI groups was similar to that in the control group (8-10%) which was injected with Ca2+ and polyvinyl pyrrolidone only. The proportion of blastocysts that developed in culture from the ICSI-derived 2-cell embryos was of the order of 36-42%. Some blastocysts were used for cell number counts. There was a significant increase in total and inner cell mass counts of blastocysts in which the spermatozoon was injected at 2 and 3 h following Ca2+. The remaining blastocysts were transferred to day 3 pseudopregnant mice, of which 33% subsequently became pregnant. Of the blastocysts transferred, 16-25% developed to term in vivo. No deformities were observed in the pups. We believe this is the first report of live-birth following mouse ICSI.
Source Title: Human Reproduction
URI: http://scholarbank.nus.edu.sg/handle/10635/131312
ISSN: 02681161
Appears in Collections:Staff Publications

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