Culturing, Freezing, Processing, and Imaging of Entire Organoids and Spheroids While Still in a Hydrogel

作者： Olgu Enis Tok1,2, Gamze Demirel2,3, Yusuf Saatci1, Zeynep Akbulut3,5, Ozgecan Kayalar6, Ranan Gulhan Aktas2,3,4 1Research Institute for Health Sciences and Technologies (SABITA),Istanbul Medipol University, 2Cellorama, 3Cancer and Stem Cell Research Center,Maltepe University, 4Department of Histology and Embryology, School of Medicine,Maltepe University, 5Department of Medical Biology, School of Medicine,Maltepe University, 6School of Medicine,Koc University

简介：

Overview

This study addresses the challenges associated with culturing organoids and spheroids, which are delicate 3D structures. A multipurpose device is employed to maintain their integrity during various procedures, such as freezing, thawing, and staining for examination.

Key Study Components

Research Area

• Cell biology
• Organoid and spheroid culture
• Microscopic imaging techniques

Background

• Organoids and spheroids are prone to damage during manipulation.
• Maintaining their structural integrity enhances reproducibility in biological research.
• The study focuses on simplifying processes involving these 3D structures.

Methods Used

• The study entails culturing, freezing, thawing, and staining organoids and spheroids.
• HEPG2 cells were utilized as a model organism.
• A multipurpose device and various microscopy techniques were key technologies.

Main Results

• Successful maintenance of organoid and spheroid integrity during various procedures.
• Immunofluorescent labeling confirmed effective visualization of biological markers.
• The method produced consistent results across different 3D systems.

Conclusions

• The protocol effectively preserves organoids and spheroids, allowing for reliable analysis.
• This advancement has significant implications for disease modeling and biomarker examination.

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