Integration of single oocyte trapping, in vitro fertilization and embryo culture in a microwell-structured microfluidic device

Abstract

In vitro fertilization (IVF) therapy is an important treatment for human infertility. However, the methods for clinical IVF have only changed slightly over decades: culture medium is held in oil-covered drops in Petri dishes and manipulation occurs by manual pipetting. Here we report a novel microwell-structured microfluidic device that integrates single oocyte trapping, fertilization and subsequent embryo culture. A microwell array was used to capture and hold individual oocytes during the flow-through process of oocyte and sperm loading, medium substitution and debris cleaning. Different microwell depths were compared by computational modeling and flow washing experiments for their effectiveness in oocyte trapping and debris removal. Fertilization was achieved in the microfluidic devices with similar fertilization rates to standard oil-covered drops in Petri dishes. Embryos could be cultured to blastocyst stages in our devices with developmental status individually monitored and tracked. The results suggest that the microfluidic device may bring several advantages to IVF practices by simplifying oocyte handling and manipulation, allowing rapid and convenient medium changing, and enabling automated tracking of any single embryo development.

Rapid, efficient functional characterization and recovery of HIV-specific human CD8 + T cells using microengraving

“…Arrays of 1-mm-thick polydimethylsiloxane nanowells (50 μm) were manufactured by replica molding using a custombuilt mold, adhered directly to a 3 in ×1 in glass slide, and sterilized in an oxygen plasma before use (41). anti-CD28 and anti-CD49d for 5 h. Stimulated cells were counted and adjusted to a density of 1 × 10 6 cells/mL A 300-μL aliquot of the cells was deposited onto a sterilized array of nanowells.…”

Paper-Based Quantification of Male Fertility Potential

“…In the context of fertility, traditional channel-based devices have been developed for sperm selection (18 -22 ), embryo development (23)(24)(25), and semen analysis (14, 26 -28 ). Recently, paper has emerged as a scalable diagnostic platform (29 -32 ).…”

Application of microfluidic technologies to human assisted reproduction

“…Microfluidic IVF/CI in animal model studies has also emerged as a step in the process of 'LOC' IVF. A sophisticated microfluidic device was used for trapping (spatially maintaining) mouse oocytes, motile sperm isolation, microinsemination and embryo development for 96 h (Han et al, 2010). This report demonstrated that all these steps in the mouse IVF process could be performed on a single device.…”

“…In addition, an in silico modelling experiment was performed to begin explaining potential mechanisms of benefit of culturing in a microfunnel with dynamic media flow compared with microchannels with gravity flow (detrimental to embryo development; Hickman et al, 2002). Han et al (2010) also reported a microwell structure microfluidic device that performed multiple steps of the IVF process. Using mouse embryos they demonstrated that this microfluidic cartridge could trap oocytes in microwells, integrate insemination and fertilization and support embryo development to the blastocyst stage.…”