A microfluidic perfusion approach for on-chip characterization of the transport properties of human oocytes

Abstract

Accurate characterization of the cell membrane transport properties of human oocytes is of great significance to reproductive pharmacology, fertility preservation, and assisted reproduction. However, the commonly used manual method for quantifying the transport properties is associated with uncontrolled operator-to-operator and run-to-run variability. Here, we report a novel sandwich structured microfluidic device that can be readily fabricated for characterizing oocyte membrane transport properties. Owing to its capacity of excellent control of both solution replacement and temperature in the microchannel, the temperature-dependent permeability of oocyte membrane can be precisely characterized. Furthermore, the fertilization and developmental competence analysis post perfusion indicate that our approach does not compromise the physiological function of in-vitro matured human oocytes. Collectively, we present the development of a novel sandwich structured microfluidic device based approach that allows for on-chip characterization of the transport properties of human oocytes under innocuous osmotic shock or injury to the cells.

A highly-occupied, single-cell trapping microarray for determination of cell membrane permeability

“…The micropipette perfusion technique has been commonly used to determine the membrane permeability of animal and human oocytes to water and CPAs, 17, 18 which may suffer from uncontrolled operation-to-operation variability. 19 …”

“…19–22 Microfluidic devices offer great advantages such as reduced reagent volume, shortened reaction time, and parallel operation. 23 Chen et al 20 developed a microfluidic perfusion chamber to measure the volumetric change of rat basophilic leukemia cells following exposure to hypertonic solutions.…”

“…A group of cells (usually 10-15) could be trapped along the obstacle edge, but they might interact with each other in an uncontrollable way, since the device was not intrinsically designed to keep trapped cells apart. Similarly, Zhao et al 19 designed a perfusion chamber based on the same principle of “road blocking” and further integrated it with a temperature-controlled stage to specifically measure the osmotic behavior of human oocytes at 4-25°C. The proposed device was not made of polydimethylsiloxane (PDMS), and therefore the fabrication did not require soft lithography and plasma treatment.…”

Surface‐Acoustic‐Wave‐Based Lab‐on‐Chip for Rapid Transport of Cryoprotectants across Cell Membrane for Cryopreservation with Significantly Improved Cell Viability

“…As the CPA loading and unloading processes and methods are fundamental and extremely important for cryopreservation, great efforts have been devoted for the development of new and advanced methods and process for them. Researchers studied human umbilical vein endothelial cells (HUVECs) in a self‐made microperfusion chamber, measured their cryobiological properties, and optimized the removal of 1.5 m DMSO at 25 °C . Benson et al and Davidson et al optimized cryoprotectant equilibration using a piecewise‐constant procedure, reduced toxicity, and osmotic shock for oocyte cryopreservation .…”

A Multiwell Microfluidic Device for Analyzing and Screening Nonhormonal Contraceptive Agents

“…Cell‐based drug screening platforms that expose target cells with individual drugs include microreactors, droplets, and hydrogels . To maintain the cells during biochemical procedures (e.g., washing), engineering techniques, including geometrical design, microarrays, membrane, and dielectrophoresis, have been incorporated into drug screening systems. However, these techniques do not address the requirements for the analysis of COC development that requires multiple enzyme loading, cumulus cell removal, and straining procedures .…”