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Gene Transfection toward Spheroid Cells on Micropatterned Culture Plates for Genetically-modified Cell Transplantation

作者： Keiji Itaka1, Satoshi Uchida1, Akitsugu Matsui1, Kayoko Yanagihara1, Masaru Ikegami1, Taisuke Endo2, Takehiko Ishii3, Kazunori Kataoka2 1Graduate School of Medicine, Laboratory of Clinical Biotechnology,The University of Tokyo, 2Graduate School of Engineering, Department of Materials Engineering,The University of Tokyo, 3Graduate School of Engineering, Department of Bioengineering,The University of Tokyo

简介：

Overview

This protocol describes a cell transplantation system using genetically modified, injectable spheroids. The system facilitates prolonged transgene expression from the transplanted cells in host animals while maintaining the innate function of the cells.

Key Study Components

Area of Science

Cell transplantation
Genetic modification
3D cell culture

Background

Cell spheroids are cultured on micropatterned culture plates.
Gene introduction is achieved using polyplex nanomicelles.
This method enhances the functionality of transplanted cells.
Maintaining innate cell function is crucial for successful transplantation.

Purpose of Study

To develop a method for effective cell transplantation.
To ensure prolonged expression of transgenes in host animals.
To maintain the innate function of genetically modified cells.

Methods Used

Preparation of 3D cell spheroids using rat hepatocytes or MSCs.
Mixing polymers and plasmid DNA to create polyplex nanomicelles.
Transfection of cells with the polyplex solution.
Recovery of spheroids for transplantation.

Main Results

Successful recovery of genetically modified spheroids.
Prolonged transgene expression observed in host animals.
Innate function of cells maintained post-transplantation.
Demonstrated by laboratory personnel.

Conclusions

The protocol provides a reliable method for cell transplantation.
Genetic modification enhances the therapeutic potential of transplanted cells.
Maintaining cell function is essential for successful outcomes.

Frequently Asked Questions

What types of cells can be used for spheroid formation?

Rat hepatocytes or mesenchymal stem cells (MSCs) can be used.

How are the spheroids recovered after gene introduction?

Spheroids are recovered by placing the plates on ice and aspirating the detached spheroids.

What is the role of polyplex nanomicelles in this protocol?

They are used for gene introduction into the cells.

What is the significance of maintaining innate cell function?

It is crucial for the therapeutic efficacy of the transplanted cells.

Who demonstrated the procedures in the study?

The procedures were demonstrated by laboratory personnel including Ms. Ula, Ms. Yha, Ms. Suzuki, Mr. Matsui, and Dr. They.

This protocol describes a cell transplantation system using genetically modified, injectable spheroids. Cell spheroids are cultured on micropatterned culture plates and recovered after gene introduction using polyplex nanomicelles. This system facilitates prolonged transgene expression from the transplanted cells in host animals while maintaining the innate function of the cells.