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A modified in vitro tool for isolation and characterization of rat quiescent islet stellate cells
Southeast Univ, Sch Med, Inst Diabet, Zhongda Hosp,Dept Endocrinol, Nanjing, Jiangsu, Peoples R China.
Southeast Univ, Sch Med, Inst Diabet, Zhongda Hosp,Dept Endocrinol, Nanjing, Jiangsu, Peoples R China.
Southeast Univ, Sch Med, Inst Diabet, Zhongda Hosp,Dept Endocrinol, Nanjing, Jiangsu, Peoples R China.
Southeast Univ, Sch Med, Inst Diabet, Zhongda Hosp,Dept Endocrinol, Nanjing, Jiangsu, Peoples R China.
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2019 (English)In: Experimental Cell Research, ISSN 0014-4827, E-ISSN 1090-2422, Vol. 384, no 1, article id 111617Article in journal (Refereed) Published
Abstract [en]

Background: Islet stellate cells (ISCs) play a critical role in islet fibrosis, contributing to the progression of pancreatic diseases. Previous studies have focused on fibrosis-associated activated ISCs obtained by standard islet explant techniques. However, in vitro models of quiescent ISCs (qISCs) are lacking. This study aims to develop a method to isolate qISCs and analyze their phenotype during activation.

Methods: Immunofluorescence staining was applied to localize ISCs in normal human, rat, and mouse islets. qISCs were isolated from rat islets using density gradient centrifugation (DGC) method. qRT-PCR, immunoblotting, proliferation, and migration assays were employed for their characterization.

Results: Desmin-positive ISCs were detected in normal human, rat, and mouse islets. Freshly isolated qISCs, obtained by density gradient centrifugation, displayed a polygonal appearance with refringent cytoplasmic lipid droplets and expressed transcriptional markers indicating a low activation/quiescent state. With increasing culture time, the marker expression pattern changed, reflecting ISC activation. qISCs contained more lipid droplets and exhibited lower proliferation and migration abilities compared to spindle-shaped ISCs obtained by traditional explant techniques.

Conclusions: This study describes a new method for efficient isolation of qISCs from rat islets, representing a useful in vitro tool to study the biology of ISCs in more physiological conditions.

Place, publisher, year, edition, pages
ELSEVIER INC , 2019. Vol. 384, no 1, article id 111617
Keywords [en]
Islet stellate cell, Isolation, Quiescent, Activated, Method
National Category
Cell and Molecular Biology Endocrinology and Diabetes
Identifiers
URN: urn:nbn:se:uu:diva-396432DOI: 10.1016/j.yexcr.2019.111617ISI: 000490031700012PubMedID: 31505166OAI: oai:DiVA.org:uu-396432DiVA, id: diva2:1368106
Available from: 2019-11-06 Created: 2019-11-06 Last updated: 2019-11-06Bibliographically approved

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Carlsson, Per-Ola

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