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Fluorescein-based sensors to purify human α-cells for functional and transcriptomic analyses.

ABSTRACT: Pancreatic α-cells secrete glucagon, an insulin counter-regulatory peptide hormone critical for the maintenance of glucose homeostasis. Investigation of the function of human α-cells remains a challenge due to the lack of cost-effective purification methods to isolate high-quality α-cells from islets. Here, we use the reaction-based probe diacetylated Zinpyr1 (DA-ZP1) to introduce a novel and simple method for enriching live α-cells from dissociated human islet cells with ~95% purity. The α-cells, confirmed by sorting and immunostaining for glucagon, were cultured up to 10 days to form α-pseudoislets. The α-pseudoislets could be maintained in culture without significant loss of viability, and responded to glucose challenge by secreting appropriate levels of glucagon. RNA-sequencing analyses (RNA-seq) revealed that expression levels of key α-cell identity genes were sustained in culture while some of the genes such as DLK1, GSN, SMIM24 were altered in α-pseudoislets in a time-dependent manner. In conclusion, we report a method to sort human primary α-cells with high purity that can be used for downstream analyses such as functional and transcriptional studies.

SUBMITTER: Kahraman S

PROVIDER: S-EPMC10567109 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature

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Fluorescein-based sensors to purify human α-cells for functional and transcriptomic analyses.

Kahraman Sevim S Shibue Kimitaka K De Jesus Dario F DF Kim Hyunki H Hu Jiang J Manna Debasish D Wagner Bridget B Choudhary Amit A Kulkarni Rohit N RN

eLife 20230921

Pancreatic α-cells secrete glucagon, an insulin counter-regulatory peptide hormone critical for the maintenance of glucose homeostasis. Investigation of the function of human α-cells remains a challenge due to the lack of cost-effective purification methods to isolate high-quality α-cells from islets. Here, we use the reaction-based probe diacetylated Zinpyr1 (DA-ZP1) to introduce a novel and simple method for enriching live α-cells from dissociated human islet cells with ~95% purity. The α-cell ...[more]

PMID: 37732504

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Dataset Information

Fluorescein-based sensors to purify human α-cells for functional and transcriptomic analyses.

Publications

Fluorescein-based sensors to purify human α-cells for functional and transcriptomic analyses.

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