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体外诱导大鼠骨髓间质干细胞分化为胰岛素分泌细胞

贾延劼 钟乐 宋建辉 罗芳 孙吉平 杨于嘉

中国当代儿科杂志2003,Vol.5Issue(5):393-397,5.
中国当代儿科杂志2003,Vol.5Issue(5):393-397,5.

体外诱导大鼠骨髓间质干细胞分化为胰岛素分泌细胞

Rat Bone Marrow Mesenchymal Stem Cells Transdifferentiate into Islet-secreting Cells in Vitro

贾延劼 1钟乐 1宋建辉 1罗芳 1孙吉平 1杨于嘉1

作者信息

  • 1. 中南大学湘雅医院儿科,湖南,长沙,410008
  • 折叠

摘要

Abstract

Objective To explore the protocol that induced marrow mesenchymal stem cells (MSCs) differentiating into islet-secreting cells in vitro and to provide new clues for the sources of islet transplantation.Methods Using a defined culture medium and technique for transdifferentiation, MSCs from adult SD rats were guided into specific insulin-secreting cells. The expressions of nestin and islet-specific hormones and proteins, such as insulin, glucagon, somatostatin and pancreatic duodenal homeobox 1 (Pdx-1) were analyzed by indirect immunofluorescence cytochemistry staining before and after induction. The expressions of pancreatic islet cell differentiation-related transcripts, such as nestin, insulin 1, glucose transporter 2 (GLUT 2), Isl-1, Pdx-1, Pax-4 and Pax-6 were detected by reverse transcription-PCR (RT-PCR). In addition, the quantity of insulin secretion was examined using radioimmunoassay. Results Five hours after induction, (44.6±7.3)% of differentiated MSCs expressed nestin and it increased to (61.8±8.4)% 24 hs after induction, but the expression of nestin almost disappeared at day 14. In the meantime, islet-like cellular clusters appeared after day 14 and became more apparent by day 28. Differentiated cells were found to be immunoreactive to insulin, glucagon, somatostatin and Pdx-1, and expressed insulin 1, GLUT 2, GK, Isl-1, PDX-1, Pax-4, Pax-6 mRNA. In addition, the results of cumulative quantities of insulin of 24 hs and the stimulation index showed that differentiated cells were able to produce insulin at higher levels, and displayed glucose-dependent insulin release in vitro. Conclusions Adult rat MSCs can be differentiated into insulin-secreting cells in vitro. This approach might lead to widespread cell replacement therapy for Type 1 diabetes.

关键词

骨髓间质干细胞/胰岛/糖尿病/横向分化

Key words

Marrow mesenchymal stem cells/Islet of Langerhans/Diabetes mellitus/Transdifferentiation

分类

医药卫生

引用本文复制引用

贾延劼,钟乐,宋建辉,罗芳,孙吉平,杨于嘉..体外诱导大鼠骨髓间质干细胞分化为胰岛素分泌细胞[J].中国当代儿科杂志,2003,5(5):393-397,5.

基金项目

National Natural Science Foundation (No.30200128) (No.30200128)

中国当代儿科杂志

OACSTPCD

1008-8830

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