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Cryobanking of Human Distal Lung Epithelial Cells for Preservation of Their Phenotypic and Functional Characteristics. | LitMetric

AI Article Synopsis

  • The study focuses on maintaining the epithelium in human lungs using regional stem cells, specifically basal cells and alveolar type 2 (AT2) pneumocytes; however, methods for preserving AT2 cells effectively have been inadequate.* -
  • The researchers developed a cryobanking technique for both mouse and human lung epithelial cells, ensuring that their functional characteristics remain intact for future research.* -
  • Results showed that cryobanked AT2 cells retained their important markers and functions similar to freshly dissociated cells, making this technique a valuable resource for studying lung cell behavior and molecular features.*

Article Abstract

The epithelium lining airspaces of the human lung is maintained by regional stem cells, including basal cells of pseudostratified airways and alveolar type 2 (AT2) pneumocytes of the gas-exchange region. Despite effective techniques for long-term preservation of airway basal cells, procedures for efficient preservation of functional epithelial cell types of the distal gas-exchange region are lacking. Here we detail a method for cryobanking of epithelial cells from either mouse or human lung tissue for preservation of their phenotypic and functional characteristics. Flow cytometric profiling, epithelial organoid-forming efficiency, and single-cell transcriptomic analysis were used to compare cells recovered from cryobanked tissue with those of freshly dissociated tissue. AT2 cells within single-cell suspensions of enzymatically digested cryobanked distal lung tissue retained expression of the pan-epithelial marker CD326 and the AT2 cell surface antigen recognized by monoclonal antibody HT II-280, allowing antibody-mediated enrichment and downstream analysis. Isolated AT2 cells from cryobanked tissue were comparable with those of freshly dissociated tissue both in their single-cell transcriptome and their capacity for organoid formation in three-dimensional cultures. We conclude that the cryobanking method described herein allows long-term preservation of distal human lung tissue for downstream analysis of lung cell function and molecular phenotype and is ideally suited for the creation of an easily accessible tissue resource for the research community.

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Source
http://dx.doi.org/10.1165/rcmb.2021-0507MADOI Listing

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