Optimized Gene Transduction in Human Lung Organoids: A High-efficiency Method for Advanced Research Applications

Overview

Journal Commun Biol

Date 2025 Feb 3

PMID 39900972

Authors

Jasmin Khateeb

Jady Liang

Yuchong Li

Thenuka Thanabalasingam

Julie Khang

Mirjana Jerkic

Giovanna Pellecchia

Bhooma Thiruv

Ya-Wen Chen

Ori Rotstein

Arthur S Slutsky

Haibo Zhang

Affiliations

Soon will be listed here.

Abstract

Human induced pluripotent stem cell (iPSC)-derived lung organoids, engineered to carry targeted genes, offer a robust platform for investigating mechanistic insights in lung research. Although lentiviral vectors (LVVs) are highly effective for stable expression due to their integrative properties, achieving efficient transduction in human iPSC-derived lung organoids poses a significant technical challenge, likely due to the complex structure of these organoids. In this study, we optimized a method to enhance LVV transduction efficiency by physically disrupting the organoids to increase surface area, followed by spinoculation to apply shear force during cell dissociation. This approach, combined with the use of an optimized culture medium, significantly improved transduction efficiency. The success of this method was validated at both the gene and protein levels using single-cell RNA sequencing (scRNA-seq) and various cellular and molecular assays. Our optimized transduction protocol may provide a valuable tool for investigating specific cellular and molecular mechanisms in development and disease models using human iPSCs-derived lung organoids.

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