Defined conditions for long-term expansion of murine and human alveolar epithelial stem cells in three-dimensional cultures

Satoshi Konishi; Aleksandra Tata; Purushothama Rao Tata

doi:10.1016/j.xpro.2022.101447

Defined conditions for long-term expansion of murine and human alveolar epithelial stem cells in three-dimensional cultures

STAR Protoc. 2022 Jun 10;3(2):101447. doi: 10.1016/j.xpro.2022.101447. eCollection 2022 Jun 17.

Authors

Satoshi Konishi¹, Aleksandra Tata¹, Purushothama Rao Tata^{1

2

3

4

5}

Affiliations

¹ Department of Cell Biology, Duke University School of Medicine, Durham, NC 27710, USA.
² Division of Pulmonary Critical Care, Department of Medicine, Duke University School of Medicine, Durham, NC 27710, USA.
³ Duke Cancer Institute, Duke University School of Medicine, Durham, NC 27710, USA.
⁴ Center for Advanced Genomic Technologies, Duke University, Durham, NC 27710, USA.
⁵ Duke Regeneration Center, Duke University School of Medicine, Durham, NC 27710, USA.

Abstract

Alveolar type 2 cells (AT2s) serve as stem cells of the alveoli and restore cell numbers after injury. Here, we describe a detailed protocol for the isolation, purification, and culture of murine and human AT2s. We have developed chemically defined and stroma-free culture conditions that enable expansion and maintenance of AT2s. The culture conditions are scalable and compatible with high-throughput chemical and genetic screenings and can potentially be used to generate large AT2 numbers for cell-based therapies. For complete details on the use and execution of this protocol, please refer to Katsura et al. (2020).

Keywords: Cell Biology; Cell culture; Cell isolation; Organoids; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Alveolar Epithelial Cells*
Animals
Cell Differentiation / genetics
Humans
Lung
Mice
Pulmonary Alveoli*
Stem Cells

Abstract

Publication types

MeSH terms

Grants and funding