Abstract
Studies have demonstrated that mesenchymal stem-like cells can be isolated from endometrium. However, the potential of endometrial-derived stem cells to differentiate into insulin-positive cells and functionally secrete insulin remains undetermined. We isolated endometrial mesenchymal stem-like cells (EMSCs) from human endometrial tissue from six donors. The insulin-secreting function of EMSCs was further analyzed in vitro and in transplanted grafts in vivo. We successfully isolated EMSCs from human endometrium, and our results showed that EMSCs expressed high levels of stemness genes (Nanog, Oct-4, Nestin). Under specific induction conditions for 2 weeks, EMSCs formed three-dimensional spheroid bodies (SBs) and secreted C-peptide. The high insulin content of SB-EMSCs was confirmed by enzyme-linked immunosorbent assay, and glucose responsiveness was demonstrated by measuring glucose-dependent insulin secretion. Using cDNA microarrays, we found that the expression profiles of SB-EMSCs are related to those of islet tissues. Insulin and C-peptide production in response to glucose was significantly higher in SB-EMSCs than in undifferentiated EMSC controls. Furthermore, upon differentiation, SB-EMSCs displayed increased mRNA expression levels of NKx2.2, Glut2, insulin, glucagon, and somatostatin. Our results also showed that SB-EMSCs were more resistant to oxidative damage and oxidative damage-induced apoptosis than fibroblasts from the same patient. It is noteworthy that SB-EMSCs xenotransplanted into immunocompromised mice with streptozotocin-induced diabetes restored blood insulin levels to control values and greatly prolonged the survival of graft cells. These data suggest that EMSCs not only play a novel role in the differentiation of pancreatic progenitors, but also can functionally enhance insulin production to restore the regulation of blood glucose levels in an in vivo transplantation model.
Footnotes
This study was supported by the National Science Council of the Republic of China [Grants 97-3111-B-075-001-MY3, 97-2320-B-075-003-MY3], Taipei Veterans General Hospital [Grants V97B-004,V97B1-006, V98A-093, V99C1-166, V99A-065, E1-008, F-001, B99A-065], the Joint Projects of the Veterans General Hospitals University System of Taiwan [Grant VGHUST 98-G6-6], Yen-Tjing-Ling Medical Foundation [Grants 95/96/97/98, CI-97-6, CI-99-7], National Yang-Ming University (Ministry of Education, Aim for the Top University Plan) and Genomic Center Project, Technology Development Program for Academia, Department of Industrial Technology, Ministry of Economic Affairs [Grant 98-EC-17-A-19-S2-0107], and the Center of Excellence for Cancer Research at Taipei Veterans General Hospital [Grant DOH99-TD-C-111-007].
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
doi:10.1124/jpet.110.169284.
↵ The online version of this article (available at http://jpet.aspetjournals.org) contains supplemental material.
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ABBREVIATIONS:
- EMSC
- endometrial mesenchymal stem-like cell
- BMSC
- bone marrow-derived mesenchymal stem cell
- AMSC
- adipose tissue-derived mesenchymal stem cell
- SB
- spheroid-like body
- Q-PCR
- quantitative reverse transcription-polymerase chain reaction
- DMEM
- Dulbecco's modified Eagle's medium
- DMEM-LG
- DMEM with 1 g/l of glucose
- FBS
- fetal bovine serum
- SCID
- severe combined immunodeficiency
- H&E
- hemoatoxylin and eosin
- DAPI
- 4,6-diamidino-2-phenylindole
- 3D
- three-dimensional
- STZ
- streptozotocin
- PBS
- phosphate-buffered saline
- FITC
- fluorescein isothiocyanate
- MTT
- methyl thiazol tetrazolium
- IL-1β
- interleukin-1β
- GSH
- glutathione
- ROS
- reactive oxygen species
- EGF
- epidermal growth factor
- bFGF
- basic fibroblast growth factor
- PPARγ2
- peroxisome proliferator-activated receptor γ2
- MAP-2
- microtubule-associated protein 2.
- Received April 24, 2010.
- Accepted September 17, 2010.
- Copyright © 2010 by The American Society for Pharmacology and Experimental Therapeutics
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