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Hum. Reprod. Advance Access published online on September 30, 2005
Human Reproduction, doi:10.1093/humrep/dei323
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received March 20, 2005
Revised July 25, 2005
Accepted August 17, 2005

Article

IVF within microfluidic channels requires lower total numbers and lower concentrations of sperm

Ronald S. Suh 1, Xiaoyue Zhu 2, Nandita Phadke 2, Dana A. Ohl 1, Shuichi Takayama 3, and Gary D. Smith 4*

1 Department of Urology, University of Michigan, Ann Arbor, MI 48109, USA
2 Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA
3 Department of Biomedical Engineering, Department of Macromolecular Science and Engineering, University of Michigan, Ann Arbor, MI 48109, USA
4 Department of Urology, Department of Obstetrics and Gynecology, Department of Molecular and Integrated Physiology, University of Michigan, Ann Arbor, MI 48109, USA

* To whom correspondence should be addressed.
Gary D. Smith, E-mail: smithgd{at}med.umich.edu


  Abstract

BACKGROUND: Microfluidic technology has been utilized in numerous biological applications specifically for miniaturization and simplification of laboratory techniques. We sought to apply microfluidic technology to murine IVF. METHODS: Microfluidic devices measuring 500 µm wide, 180 µm deep, and 2.25 cm in length were designed and fabricated using poly(dimethylsiloxane) (PDMS). Controls were standard centre-well culture dishes with 500 µl of media, half of which also contained PDMS as a material control. Denuded mouse oocytes were placed into microchannels or centre-well dish controls in groups of 10, then co-incubated overnight with epididymal mouse sperm at various concentrations. Fertilization was assessed and Fisher’s exact test was used for statistical analysis (P < 0.05 significant). RESULTS: Fertilization rates between the two control groups (42%, no PDMS; 41%, with PDMS; not significant) were similar. Fertilization rates for denuded oocytes at standard mouse insemination sperm concentration (1x106 sperm/ml) was poorer in microchannels (12%) than controls (43%; P < 0.001). As insemination concentrations decreased, fertilization rates improved in microchannels with a plateau between 8x104 and 2x104 sperm/ml (4000-1000 total sperm). At these concentrations, combined fertilization rate for denuded oocytes was significantly higher in microchannels than centre-well dishes (27 versus 10%, respectively; P < 0.001), and was not significantly different from corresponding controls with a sperm concentration of 1x106 (37%; P = 0.06). CONCLUSIONS: Murine IVF can be conducted successfully within microfluidic channels. Lower total numbers and concentrations of sperm are required. Microfluidic devices may ultimately be useful in clinical IVF.

Keywords: IVF/microfluidics/murine/sperm.
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