In Vitro Culture of Single Bovine Embryos with Microwell Plates Made of Poly(dimethylsiloxane) Cured under Low Pressure

Abstract

Single embryo culture is useful for assessing the developmental competence of an embryo in detail. Recently, a device made of poly(dimethylsiloxane) (PDMS), which is biocompatible and nontoxic, has been widely used for culture various types of cells. However, PDMS plates are porous, causing the serious osmolality increment of the medium (over 600 mOsm/kg from Day 4 to Day 7). Here, we report that curing the PDMS under low pressure (LP-PDMS) greatly reduced the porosity, resulting in a constant osmolality of the medium. The blastocyst rate of single bovine embryos cultured with LP-PDMS microwell (MW) plates was the same as that of group-cultured embryos (25 embryos/50 μl droplet; control, P>0.05). These results indicate that MWs on a plate made of PDMS cured under low pressure can be successfully used for individual embryo culture.

Surface functionalization of poly(dimethylsiloxane) substrates facilitates culture of pre-implantation mouse embryos by blocking non-selective adsorption

“…All rights reserved. [22]. While PDMS substrates, when functionalized with extracellular matrix molecules, support the in vitro culture of various cell types, untreated, native PDMS cannot support embryo culture even though adhesive signalling is not required.…”

“…The majority of these devices are made from poly(dimethylsiloxane) (PDMS), which is one of the most ubiquitous polymers used in biological applications and offers straightforward prototyping [12], gas permeability [13], general biocompatibility [14,15] and optical transparency [16]. However, especially at the small fluid volumes typically used in the embryo culture, PDMS substrates have two well-documented issues: small molecule absorption/adsorption [17–21] and evaporation-mediated osmolality shifts [22]. While PDMS substrates, when functionalized with extracellular matrix molecules, support the in vitro culture of various cell types, untreated, native PDMS cannot support embryo culture even though adhesive signalling is not required.…”