Liquid Marble Microbioreactor System: A 3D Culture Protocol for Encapsulation of Porcine Oocytes

Abstract

Source: Gorczyca, G., et al. Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation. J. Vis. Exp. (2020).

In this video, we describe a protocol for the encapsulation of porcine oocytes in hydrophobic microbioreactors.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Encapsulation in super-hydrophobic fluorinated ethylene propylene (FEP) microbioreactors.

NOTE: Make sure that all in vitro maturation (IVM) procedures are carried out on a thermostatically controlled table, and cumulus-oocyte complexes (COCs) are maintained at 38 °C throughout their handling.

1. Prepare a 30 mm Petri dish containing 5 g of FEP powder bed-average particle size of 1 μm
   NOTE: It is important to....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Maturation Medium
hCG (1 VIAL of 10 000 U)	Sigma-Aldrich	CG10
PMSG	BioVendor	RP1782721000
DMEM/F12 (500ml)	Sigma-Aldrich	D8062	Maturation Medium
FCS (100 ml)	Thermo Fisher	16140063	10% final concentration for Maturation Medium
Liquid Marble
FEP	Dyneon GmbH 3M AdMD	A-66670
Specific instruments, tools
30 mm Petri dish	TPP	93040
60 mm IVF Petri dish	Falcon	353653
Ez-Grid Premium Cell Handling Pipettor	RI Life Sciences	8-72-288
Ez-Tip	RI Life Sciences	8-72-4155/20
Incubator	Panasonic	MCO-170AIC-PE	Other brands can be used
Sterile petri dish (10 cm)	NEST Biotechnology	704002