This proposal is a design-directed project to develop a novel microfluidic bioreactor that will culture multiple single embryos under simulated physiological conditions while simultaneously performing real-time monitoring of biochemical markers of embryo quality. Reducing the incidence of high-order multiple pregnancies while maintaining the overall IVF success Irate is a holy grail of human IVF and would be greatly assisted by the ability to produce and identify the highest quality embryos. This goal has been elusive to date due, at least in part, to the lack of instrumentation to perform convenient and reliable single embryo manipulation and analysis. Because of the small quantities of biomarkers produced by single embryos, reliable quantification hinges on the ability to culture the embryos in very small volumes of fluid and to directly analyze the culture media for soluble biomarkers secreted by embryos with minimal dilution. The proposed microbioreactor with microfluidic pumps, valves, and sensors will provide an inherently biomimetic milieu for embryo culture as well as enable direct biomarker analysis on chip. The microbioreactor will utilize a computer-controlled integrated microfluidics platform that controls fluid flow inside elastomeric capillaries by deformation of the microchannels with mechanical microactuators. The biomarkers of mouse embryo health that will be monitored in this exploratory grant are embryo metabolites, autocrine factors, and embryo surface biomarker of embryo health. The embryo microbioreactor will be used to specifically test the hypothesis that analysis of select biochemical markers will enable prediction of which 8 cell embryo will proceed to produce healthy blastocysts. This test will mainly serve the purpose of device validation and concept feasibility but it also has clinical relevance. A current trend is to grow embryos to the blastocyst stage and transfer the two morphologically """"""""best"""""""" blastocyst. Recently, however, two reports using mouse embryos have independently demonstrated that extended culture to the blastocyst stage causes aberrant genetic imprinting and altered postnatal development, growth, physiology and behavior. Case reports and studies also suggest a general association between in vitro culture to the blastocyst stage and MZ twinning. This proposal will address these issues by developing novel non-invasive means of selecting embryos with the greatest implantation potential, with the least amount of manipulation and culture.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HD049607-02
Application #
7038279
Study Section
Special Emphasis Panel (ZRG1-ISD (01))
Program Officer
Tasca, Richard J
Project Start
2005-04-01
Project End
2008-03-31
Budget Start
2006-04-01
Budget End
2008-03-31
Support Year
2
Fiscal Year
2006
Total Cost
$160,396
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Biomedical Engineering
Type
Schools of Engineering
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
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Smith, Gary D; Monteiro da Rocha, Andre (2012) Advances in embryo culture systems. Semin Reprod Med 30:214-21
Heo, Yun Seok; Cabrera, Lourdes M; Bormann, Charles L et al. (2012) Real time culture and analysis of embryo metabolism using a microfluidic device with deformation based actuation. Lab Chip 12:2240-6
Smith, G D; Swain, J E; Bormann, C L (2011) Microfluidics for gametes, embryos, and embryonic stem cells. Semin Reprod Med 29:5-14
Heo, Y S; Cabrera, L M; Bormann, C L et al. (2010) Dynamic microfunnel culture enhances mouse embryo development and pregnancy rates. Hum Reprod 25:613-22
Smith, G D; Takayama, S (2007) Gamete and embryo isolation and culture with microfluidics. Theriogenology 68 Suppl 1:S190-5
Heo, Yun Seok; Cabrera, Lourdes M; Song, Jonathan W et al. (2007) Characterization and resolution of evaporation-mediated osmolality shifts that constrain microfluidic cell culture in poly(dimethylsiloxane) devices. Anal Chem 79:1126-34
Suh, Ronald S; Zhu, Xiaoyue; Phadke, Nandita et al. (2006) IVF within microfluidic channels requires lower total numbers and lower concentrations of sperm. Hum Reprod 21:477-83