Isolation of high-yield and -quality RNA from human precision-cut lung slices for RNA-sequencing and computational integration with larger patient cohorts

نویسندگان

چکیده

Precision-cut lung slices (PCLS) have gained increasing interest as a model to study biology/disease and screening novel therapeutics. In particular, PCLS derived from human tissue can better recapitulate some aspects of compared with animal models. Several experimental readouts been established for use PCLS, but obtaining high-yield -quality RNA downstream analysis has remained challenging. This is particularly problematic utilizing the power next-generation sequencing techniques, such RNA-sequencing (RNA-seq), nonbiased high-throughput cohorts. current study, we present approach isolating high-quality small amount tissue, including diseased idiopathic pulmonary fibrosis. We show that isolated using this method sufficient quality RT-qPCR RNA-seq analysis. Furthermore, data could be used in several computational pipelines, deconvolution bulk publicly available single-cell data. Deconvolution Bisque revealed diversity cell populations immune populations, which correlated known aberrant disease.

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ژورنال

عنوان ژورنال: American Journal of Physiology-lung Cellular and Molecular Physiology

سال: 2021

ISSN: ['1522-1504', '1040-0605']

DOI: https://doi.org/10.1152/ajplung.00401.2020