Isolation and Enrichment of Human Lung Epithelial Progenitor Cells for Organoid Culture

Bindu Konda; Apoorva Mulay; Changfu Yao; Stephen Beil; Edo Israely; Barry  R. Stripp

doi:10.3791/61541

Isolation and Enrichment of Human Lung Epithelial Progenitor Cells for Organoid Culture

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Biology

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Isolation and Enrichment of Human Lung Epithelial Progenitor Cells for Organoid Culture

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11:49 min

July 21, 2020

DOI: 10.3791/61541-v

Bindu Konda¹, Apoorva Mulay¹, Changfu Yao¹, Stephen Beil¹, Edo Israely¹, Barry R. Stripp¹

¹Lung and Regenerative Medicine Institutes, Department of Medicine,Cedars-Sinai Medical Center

Overview

This article outlines a comprehensive methodology for tissue dissociation and cellular fractionation techniques aimed at enriching viable epithelial cells from various regions of the human lung. The established protocols facilitate the functional analysis of lung epithelial progenitor cells through 3D organoid culture models.

Key Study Components

Research Area

Tissue dissociation and cellular fractionation
Proximal and distal lung epithelial cell enrichment
3D organoid culture models

Background

Importance of understanding lung tissue maintenance
Need for models to study lung disease and remodeling
Potential for drug discovery and validation platforms

Methods Used

Minced lung tissue digestion and isolation procedures
Human lung tissue as the biological system
Fluorescence-activated cell sorting (FACS) for cell analysis

Main Results

Successful isolation of epithelial progenitor cells from lung regions
Preservation of positional identity of isolated cells in vitro
Modeling regional differences in lung response to stimuli

Conclusions

This study provides essential protocols for lung epithelial cell enrichment.
The methodologies have significant implications for research on lung biology and disease.

Frequently Asked Questions

What types of cells are isolated using this protocol?

The protocol focuses on isolating viable epithelial progenitor cells from both proximal and distal regions of the human lung.

How does this study contribute to drug discovery?

By providing a consistent platform for modeling lung responses, the study aids in screening drugs for their efficacy against lung diseases.

What is the significance of using 3D organoid cultures?

3D organoid cultures mimic in vivo conditions better, allowing for more accurate studies of lung biology and disease mechanisms.

Can the techniques be combined with other methods?

Yes, the tissue dissociation methods are compatible with various cellular fractionation approaches.

What biological applications arise from the study?

The enriched cells can be utilized for studying fundamental lung biology, tissue maintenance, and regional responses to stimuli.

Are specific lung regions targeted in this research?

The research specifically focuses on the proximal airways and distal lung epithelium to study regional responses.

What challenges does this protocol address?

The protocol addresses challenges in isolating viable cells while maintaining their functional characteristics.

This article provides a detailed methodology for tissue dissociation and cellular fractionation approaches allowing enrichment of viable epithelial cells from proximal and distal regions of the human lung. Herein these approaches are applied for the functional analysis of lung epithelial progenitor cells through the use of 3D organoids culture models.

3D organoid cultures are proximal in distal lung epithelium developed using this protocol. Provide tractable model to study fundamental mechanisms of lung tissue maintenance or disease associated remodeling, and serves an effective platform for drug discovery and validation. This protocol is compatible with other tissue dissociation cellular fractionation approaches.

And since epithelial progenitor cells is isolated from different anatomic compartments maintain their positional identity in vitro, it can be used for modeling regional differences in lung responses to either exogenous or endogenous stimuli. This level technique can be used for the isolation of different subpopulation of epithelial cells including region specific progenitor cells that can be cultured to yield specialist differentiated progeny representative of the region of origin. Upon receive of the lung tissue, separate the proximal trachea and bronchi from the distal lung epithelium, which contains small airways of two millimeters in diameter or less, and the surrounding parenchymal tissue.

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