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在水凝胶中培养、冷冻、处理和成像整个类器官和球状体
在水凝胶中培养、冷冻、处理和成像整个类器官和球状体
JoVE Journal
Biology
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JoVE Journal Biology
Culturing, Freezing, Processing, and Imaging of Entire Organoids and Spheroids While Still in a Hydrogel

在水凝胶中培养、冷冻、处理和成像整个类器官和球状体

Full Text
8,126 Views
08:07 min
December 23, 2022

DOI: 10.3791/64563-v

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

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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.

Frequently Asked Questions

What is the significance of using a multipurpose device?
It allows for the simultaneous handling of various processes while maintaining the structural integrity of organoids and spheroids.
How does this study enhance reliability in research?
By ensuring organoids and spheroids remain intact during manipulation, it increases reproducibility and accuracy in experiments.
Can this method be applied to other biological systems?
Yes, it is adaptable to various 3D growing systems, including individual cells or different organoid types.
What are the primary challenges addressed in this study?
The study addresses the fragility of organoids and spheroids during handling and the need for reliable protocols in their study.
What types of microscopes can be used with this protocol?
The protocol is designed for use with various microscopy techniques, including inverted and confocal microscopy.
How long can organoids be stored using this method?
Organoids can be stored for longer than six months by transferring them to a liquid nitrogen tank after freezing.

本研究描述了在各种显微镜下培养、冷冻、解冻、处理、染色、标记和检查整个球状体和类器官的方法,同时它们在多用途装置内的水凝胶中保持完整。

类器官和球状体非常脆弱,在处理时会严重损坏。我们的实验方案确保这些3D生长结构在各种过程中保持完整,从而提高准确性、可重复性、可靠性和可重复性。研究人员可以在各种显微镜下培养、冷冻、解冻、处理、染色、标记和检查整个类器官和球状体,同时它们在单个多用途装置中的水凝胶中保持完整。

它可以在一个多功能设备中创建疾病模型并跟踪顺序步骤。该技术在检查用于疾病诊断和预后的生物标志物时带来了更高的准确性。该方法可应用于任何 3D 生长系统,包括类器官、球状体、单个细胞或活生物体。

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