Single-cell RNA Sequencing and Analysis of Human Pancreatic Islets.
J Vis Exp
; (149)2019 07 18.
Article
em En
| MEDLINE
| ID: mdl-31380847
ABSTRACT
Pancreatic islets comprise of endocrine cells with distinctive hormone expression patterns. The endocrine cells show functional differences in response to normal and pathological conditions. The goal of this protocol is to generate high-quality, large-scale transcriptome data of each endocrine cell type with the use of a droplet-based microfluidic single-cell RNA sequencing technology. Such data can be utilized to build the gene expression profile of each endocrine cell type in normal or specific conditions. The process requires careful handling, accurate measurement, and rigorous quality control. In this protocol, we describe detailed steps for human pancreatic islets dissociation, sequencing, and data analysis. The representative results of about 20,000 human single islet cells demonstrate the successful application of the protocol.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Ilhotas Pancreáticas
/
Análise de Sequência de RNA
/
Análise de Célula Única
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article