Rat Bone Marrow Mesenchymal Stem Cells Transdifferentiate into Islet-secreting Cells in Vitro
10.3969/j.issn.1008-8830.2003.05.001
- VernacularTitle:体外诱导大鼠骨髓间质干细胞分化为胰岛素分泌细胞
- Author:
JIA YAN-JIE
1
;
ZHONG LE
;
SONG JIAN-HUI
;
LUO FANG
;
SUN JI-PING
;
YANG YU-JIA
Author Information
1. Central South University
- Keywords:
Marrow mesenchymal stem cells;
Islet of Langerhans;
Diabetes mellitus;
Transdifferentiation
- From:
Chinese Journal of Contemporary Pediatrics
2003;5(5):393-397
- CountryChina
- Language:Chinese
-
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.
