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Rapid, Efficient, and Universally Applicable Genetic Engineering of Intestinal Organoid with a Sequential Monolayer to Three-Dimensional Strategy

Authors :
Weili Han
Guofang Lu
Sheng Zhao
Rui Wang
Haohao Zhang
Kun Liu
Yongzhan Nie
Jiaqiang Dong
Source :
Stem Cells International, Vol 2024 (2024)
Publication Year :
2024
Publisher :
Hindawi Limited, 2024.

Abstract

Genetically modified intestinal organoids are being explored as potential surrogates of immortalized cell lines and gene-engineered animals. However, genetic manipulation of intestinal organoids is time-consuming, and the efficiency is far beyond satisfactory. To ensure the yield of the genetically modified organoids, large quantity of starting materials is required, and the procedure usually takes more than 10 days. Two major obstacles that restrict the genetic delivery efficiency are the three-dimensional culture condition and that the genetic delivery is carried out in cell suspensions. In the present study, we introduce a novel highly efficient strategy for building genetically modified intestinal organoids in which genetic delivery was performed in freshly established monolayer primary intestinal epithelial cells under two-dimensional conditions and subsequentially transformed into three-dimensional organoids. The total procedure can be finished within 10 hr while displaying much higher efficiency than the traditional methods. Furthermore, this strategy allowed for the selection of transgenic cells in monolayer conditions before establishing high-purity genetically modified intestinal organoids.

Subjects

Subjects :
Internal medicine
RC31-1245

Details

Language :
English
ISSN :
16879678
Volume :
2024
Database :
Directory of Open Access Journals
Journal :
Stem Cells International
Publication Type :
Academic Journal
Accession number :
edsdoj.6185d3895469f9d0b9b05424886cf
Document Type :
article
Full Text :
https://doi.org/10.1155/2024/2005845