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ID 48976
フルテキストURL
著者
Sano, Hikaru Department of Animal Science, Graduate school of Natural Science and Technology, Okayama University
Matsuura, Koji Cardiovascular Physiology, Graduate school of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Naruse, Keiji Cardiovascular Physiology, Graduate school of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Funahashi, Hiroaki Department of Animal Science, Graduate school of Natural Science and Technology, Okayama University
抄録
The objective of this study was to use a microfluidic sperm sorter (MFSS), designed to isolate motile human spermatozoa with laminar flows (no centrifugation), for porcine IVF. Boar spermatozoa were diluted at 1 x 10(8) with a diluent containing 20% seminal fluid and flowed with modified TCM-199 (mM199, with 5 mM caffeine) to introduce motile sperm into the exit chamber for IVF. In Experiment 1, after flowing for 5 min, sperm concentration varied significantly among specific sites within the MFSS collecting chamber (range, 0.8 +/- 0.5 x 10(4) to 575.0 +/- 56.3 x 10(4) cells/mL; mean +/- SEM). In Experiment 2, when porcine IVM oocytes were placed at three locations in the MFSS exit chamber (where only motile spermatozoa accumulated) and subsequently cultured in caffeine-free mM199 for 8 h, sperm penetration rate was not significantly different among places (86.1 +/- 10.5 to 100%), but the monospermic penetration rate was lower (P < 0.05) in oocytes 3.5 mm from the exit position (12.5 +/- 4.8%) than those at 7.5 mm (53.1 +/- 6.0%) or further (41.9 +/- 2.8%) from the exit. In Experiment 3, the normal fertilization index (ratio of monospermic oocytes to number of oocytes examined) 8 h after insemination was higher (P < 0.05) in the MFSS-IVF system (0.375 +/- 0.040) than both standard IVF and transient IVF (0.222 +/- 0.028 and 0.189 +/- 0.027, respectively, with co-culture for 8 h and for 5 min). Developmental competence of fertilized oocytes (blastocyst formation) was higher (P < 0.05) in the MFSS-IVF system (40.9 +/- 2.3%) than in either standard or transient IVF (22.6 +/- 1.4 and 33.7 +/- 3.5%). In conclusion, brief co-culture of porcine oocytes with spermatozoa gradually accumulated in the MFSS chamber improved the efficiency of producing monospermic fertilized embryos and blastocysts. Furthermore, efficiencies were significantly affected by oocyte location within the chamber.
キーワード
Oocytes
Polyspermy
IVF
Sperm sorter
Pig
発行日
2010-09-15
出版物タイトル
Theriogenology
74巻
5号
出版者
Elsevier
開始ページ
863
終了ページ
870
ISSN
0093-691X
資料タイプ
学術雑誌論文
プロジェクト
異分野融合先端研究コア
オフィシャル URL
http://www.sciencedirect.com/science/article/pii/S0093691X10002013
言語
English
OAI-PMH Set
岡山大学
著作権者
Copyright © 2010 Elsevier Inc. All rights reserved.
論文のバージョン
author
査読
有り
DOI
PubMed ID
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