Apr 3, 2020

Versatile workflow for cell type resolved transcriptional and epigenetic profiles from cryopreserved human lung.

BioRxiv : the Preprint Server for Biology
M. Llamazares PradaRenata Zofia Jurkowska

Abstract

The complexity of the lung microenvironment together with changes in cellular composition during disease progression make it exceptionally hard to understand the molecular mechanisms leading to the development of chronic lung diseases. Although recent advances in cell type resolved and single-cell sequencing approaches hold great promise for studying complex diseases, their implementation greatly relies on local access to fresh tissue, as traditional methods to process and store tissue do not allow viable cell isolation. To overcome these hurdles, we developed a novel, versatile workflow that allows long-term storage of human lung tissue with high cell viability, permits thorough sample quality check before cell isolation, and is compatible with next generation sequencing-based profiling, including single-cell approaches. We demonstrate that cryopreservation is suitable for isolation of multiple cell types from different lung locations and is applicable to both healthy and diseased tissue, including COPD and tumor samples. Basal cells isolated from cryopreserved airways retain the ability to differentiate, indicating that cellular identity is not altered by cryopreservation. Importantly, using RNA sequencing (RNA-seq) and Illum...Continue Reading

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Mentioned in this Paper

Extracellular Matrix
Environment
Anatomical Space Structure
Evaluation
Location
Simulation
Species
Landscapes

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