Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation

Gabriela Gorczyca; Kamil Wartalski; Zbigniew Tabarowski; Malgorzata Duda

doi:10.3791/61325

Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte E...

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Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation

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07:45 min

July 30, 2020

DOI: 10.3791/61325-v

Gabriela Gorczyca¹, Kamil Wartalski², Zbigniew Tabarowski³, Malgorzata Duda¹

¹Department of Endocrinology, Institute of Zoology and Biomedical Research, Faculty of Biology,Jagiellonian University in Krakow, ²Department of Histology,Jagiellonian University Medical College, ³Department of Experimental Hematology, Institute of Zoology and Biomedical Research, Faculty of Biology,Jagiellonian University in Krakow

Overview

This study presents protocols for the encapsulation of porcine cumulus-oocyte complexes (COCs) under 3D culture conditions, utilizing fibrin-alginate beads and fluorinated ethylene propylene (FEP) powder. These methods maintain the spherical organization of COCs and enhance their viability for reproductive biology research and potential clinical applications.

Key Study Components

Research Area

Reproductive biology
Oocyte encapsulation
In vitro maturation methods

Background

Importance of maintaining COC structure for gap junction integrity
Potential clinical relevance for infertility treatments
Applications in biotechnology and livestock improvement

Methods Used

Encapsulation protocols using fibrin-alginate and FEP powder
Porcine oocytes as the biological system
Microscopy techniques for imaging cell viability

Main Results

Both encapsulation methods successfully maintained high viability of COCs
Granulosa cells and cumulus layers were intact in cultured COCs
Mitochondrial distribution and cell morphology were observed via transmission electron microscopy

Conclusions

The study demonstrates effective methods for encapsulating porcine oocytes to maintain viability and structure
The findings may contribute to advancements in reproductive biology and infertility therapies

Frequently Asked Questions

What are the advantages of using encapsulation for oocyte culture?

Encapsulation preserves the 3D structure of oocytes, enhancing their viability and functionality during in vitro maturation.

Can these protocols be applied to other species?

While this study focuses on porcine oocytes, similar techniques could potentially be adapted for other species with further research.

What is the role of gap junctions in oocyte health?

Gap junctions facilitate communication between oocytes and surrounding cells, crucial for nutrient exchange and overall oocyte health.

How does the choice of encapsulation material affect oocyte viability?

Different materials can influence nutrient flow and mechanical properties, which may impact how well oocytes survive and mature in culture.

What imaging techniques were used in this study?

Transmission electron microscopy was utilized to assess the ultrastructure and organelle distribution within oocytes.

Is there a clinical application for this research?

Yes, the methodologies could lead to improvements in infertility treatments by enhancing oocyte quality for IVF procedures.

What are liquid marbles, and how are they used in this study?

Liquid marbles are droplets coated with a thin layer of powder, used to encapsulate oocytes while providing a controlled microenvironment during maturation.

Presented here are two protocols for the encapsulation of porcine oocytes in 3D culture conditions. In the first, cumulus-oocyte complexes (COCs) are encapsulated in fibrin-alginate beads. In the second, they are enclosed with fluorinated ethylene propylene powder particles (microbioreactors). Both systems ensure optimal conditions to maintain their 3D organization.

This protocol for encapsulation of porcine oocytes makes it possible to maintain spherical organization of COCs. This prevents their flattening and consequent disruption of gap junctions between the oocyte and surrounding follicular cells. The main advantages of the two described protocols are that are user-friendly and allow for effective control of both the oocyte and chemo cell survival.

Both of the culture systems are valuable tools for basic research in reproductive biology, and may have clinical relevance for improved infertility treatment. They can also be applied for developing novel biotechnology methods and for livestock improvement. After rinsing the ovaries, transfer them to a beaker filled with HM medium and store them in an incubator at 38 degrees Celsius during all subsequent manipulations.

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