Investigating the suspension culture on aggregation and function of mouse pancreatic β-cells

Kai Chiang Yang, Chang Chin Wu, Shu Hua Yang, Chien Chang Chiu, Shoichiro Sumi, Hsuan Shu Lee

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The integrity and hierarchical structure of islet influence β-cells physiology dramatically. A culture substrate which can maintain or improve β-cells aggregation shall benefit cell therapy for diabetics. In this study, nontreated, type IV collagen, Lipidure, and ultralow attachment dishes were used to culture a murine β-cell line, MIN-6. The formation and biological performances of pseudoislets were investigated. Results showed that β-cells formed loose and irregular aggregates on nontreated dishes. Oppositely, pseudoislets formed on other three substrates. Most pseudoislets on Lipidure and type IV collagen dishes had a diameter between 100-150 μm with high survival rate, while large pseudoislets (>250 μm) with seriously central necrosis were found on ultralow attachment dishes. Western blot analysis revealed that pseudoislets had relatively higher connexin 36 protein productions relative to single cells. The glucose-stimulated insulin secretion test showed pseudoislets on type IV collagen have high stimulation index. Monolayers from TCPS dishes and pseudoislets from type IV collagen or Lipidure dishes were further transplanted into diabetic mice. Animals received both single cells and pseudoislets had decreasing blood glucose level and regained body weight. Histologic examination revealed that all implants successfully engrafted with positive insulin staining. Interestingly, the area under curve for the intraperitoneal glucose tolerance test showed pseudoislets had superior glucose disappearance rate. This study reveals that isolated islets or insulin-producing cells can be cultured on type IV collagen or Lipidure dishes to improve/maintain integrity prior to transplantation.

Original languageEnglish
Pages (from-to)2273-2282
Number of pages10
JournalJournal of Biomedical Materials Research - Part A
Volume101 A
Issue number8
DOIs
Publication statusPublished - Aug 2013

Fingerprint

Collagen Type IV
Collagen
Suspensions
Agglomeration
Glucose
Insulin
Physiology
Substrates
Cell culture
Blood Glucose
Monolayers
Animals
Blood
Cells
Proteins

Keywords

  • aggregation
  • pancreatic beta-cell
  • pseudoislet
  • suspension culture

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials
  • Ceramics and Composites
  • Metals and Alloys

Cite this

Investigating the suspension culture on aggregation and function of mouse pancreatic β-cells. / Yang, Kai Chiang; Wu, Chang Chin; Yang, Shu Hua; Chiu, Chien Chang; Sumi, Shoichiro; Lee, Hsuan Shu.

In: Journal of Biomedical Materials Research - Part A, Vol. 101 A, No. 8, 08.2013, p. 2273-2282.

Research output: Contribution to journalArticle

Yang, Kai Chiang ; Wu, Chang Chin ; Yang, Shu Hua ; Chiu, Chien Chang ; Sumi, Shoichiro ; Lee, Hsuan Shu. / Investigating the suspension culture on aggregation and function of mouse pancreatic β-cells. In: Journal of Biomedical Materials Research - Part A. 2013 ; Vol. 101 A, No. 8. pp. 2273-2282.
@article{2d255e79a4e146b3bb2cd470d01eb12a,
title = "Investigating the suspension culture on aggregation and function of mouse pancreatic β-cells",
abstract = "The integrity and hierarchical structure of islet influence β-cells physiology dramatically. A culture substrate which can maintain or improve β-cells aggregation shall benefit cell therapy for diabetics. In this study, nontreated, type IV collagen, Lipidure, and ultralow attachment dishes were used to culture a murine β-cell line, MIN-6. The formation and biological performances of pseudoislets were investigated. Results showed that β-cells formed loose and irregular aggregates on nontreated dishes. Oppositely, pseudoislets formed on other three substrates. Most pseudoislets on Lipidure and type IV collagen dishes had a diameter between 100-150 μm with high survival rate, while large pseudoislets (>250 μm) with seriously central necrosis were found on ultralow attachment dishes. Western blot analysis revealed that pseudoislets had relatively higher connexin 36 protein productions relative to single cells. The glucose-stimulated insulin secretion test showed pseudoislets on type IV collagen have high stimulation index. Monolayers from TCPS dishes and pseudoislets from type IV collagen or Lipidure dishes were further transplanted into diabetic mice. Animals received both single cells and pseudoislets had decreasing blood glucose level and regained body weight. Histologic examination revealed that all implants successfully engrafted with positive insulin staining. Interestingly, the area under curve for the intraperitoneal glucose tolerance test showed pseudoislets had superior glucose disappearance rate. This study reveals that isolated islets or insulin-producing cells can be cultured on type IV collagen or Lipidure dishes to improve/maintain integrity prior to transplantation.",
keywords = "aggregation, pancreatic beta-cell, pseudoislet, suspension culture",
author = "Yang, {Kai Chiang} and Wu, {Chang Chin} and Yang, {Shu Hua} and Chiu, {Chien Chang} and Shoichiro Sumi and Lee, {Hsuan Shu}",
year = "2013",
month = "8",
doi = "10.1002/jbm.a.34547",
language = "English",
volume = "101 A",
pages = "2273--2282",
journal = "Journal of Biomedical Materials Research - Part B Applied Biomaterials",
issn = "1552-4973",
publisher = "John Wiley and Sons Inc.",
number = "8",

}

TY - JOUR

T1 - Investigating the suspension culture on aggregation and function of mouse pancreatic β-cells

AU - Yang, Kai Chiang

AU - Wu, Chang Chin

AU - Yang, Shu Hua

AU - Chiu, Chien Chang

AU - Sumi, Shoichiro

AU - Lee, Hsuan Shu

PY - 2013/8

Y1 - 2013/8

N2 - The integrity and hierarchical structure of islet influence β-cells physiology dramatically. A culture substrate which can maintain or improve β-cells aggregation shall benefit cell therapy for diabetics. In this study, nontreated, type IV collagen, Lipidure, and ultralow attachment dishes were used to culture a murine β-cell line, MIN-6. The formation and biological performances of pseudoislets were investigated. Results showed that β-cells formed loose and irregular aggregates on nontreated dishes. Oppositely, pseudoislets formed on other three substrates. Most pseudoislets on Lipidure and type IV collagen dishes had a diameter between 100-150 μm with high survival rate, while large pseudoislets (>250 μm) with seriously central necrosis were found on ultralow attachment dishes. Western blot analysis revealed that pseudoislets had relatively higher connexin 36 protein productions relative to single cells. The glucose-stimulated insulin secretion test showed pseudoislets on type IV collagen have high stimulation index. Monolayers from TCPS dishes and pseudoislets from type IV collagen or Lipidure dishes were further transplanted into diabetic mice. Animals received both single cells and pseudoislets had decreasing blood glucose level and regained body weight. Histologic examination revealed that all implants successfully engrafted with positive insulin staining. Interestingly, the area under curve for the intraperitoneal glucose tolerance test showed pseudoislets had superior glucose disappearance rate. This study reveals that isolated islets or insulin-producing cells can be cultured on type IV collagen or Lipidure dishes to improve/maintain integrity prior to transplantation.

AB - The integrity and hierarchical structure of islet influence β-cells physiology dramatically. A culture substrate which can maintain or improve β-cells aggregation shall benefit cell therapy for diabetics. In this study, nontreated, type IV collagen, Lipidure, and ultralow attachment dishes were used to culture a murine β-cell line, MIN-6. The formation and biological performances of pseudoislets were investigated. Results showed that β-cells formed loose and irregular aggregates on nontreated dishes. Oppositely, pseudoislets formed on other three substrates. Most pseudoislets on Lipidure and type IV collagen dishes had a diameter between 100-150 μm with high survival rate, while large pseudoislets (>250 μm) with seriously central necrosis were found on ultralow attachment dishes. Western blot analysis revealed that pseudoislets had relatively higher connexin 36 protein productions relative to single cells. The glucose-stimulated insulin secretion test showed pseudoislets on type IV collagen have high stimulation index. Monolayers from TCPS dishes and pseudoislets from type IV collagen or Lipidure dishes were further transplanted into diabetic mice. Animals received both single cells and pseudoislets had decreasing blood glucose level and regained body weight. Histologic examination revealed that all implants successfully engrafted with positive insulin staining. Interestingly, the area under curve for the intraperitoneal glucose tolerance test showed pseudoislets had superior glucose disappearance rate. This study reveals that isolated islets or insulin-producing cells can be cultured on type IV collagen or Lipidure dishes to improve/maintain integrity prior to transplantation.

KW - aggregation

KW - pancreatic beta-cell

KW - pseudoislet

KW - suspension culture

UR - http://www.scopus.com/inward/record.url?scp=84879506251&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84879506251&partnerID=8YFLogxK

U2 - 10.1002/jbm.a.34547

DO - 10.1002/jbm.a.34547

M3 - Article

C2 - 23348877

AN - SCOPUS:84879506251

VL - 101 A

SP - 2273

EP - 2282

JO - Journal of Biomedical Materials Research - Part B Applied Biomaterials

JF - Journal of Biomedical Materials Research - Part B Applied Biomaterials

SN - 1552-4973

IS - 8

ER -