Abstract
Cellular models and culture conditions for in vitro expansion of insulin-producing cells represent a key element to develop cell therapy for diabetes. Initial evidence that human β-cells could be expanded after undergoing a reversible epithelial–mesenchymal transition has been recently negated by genetic lineage tracing studies in mice. Here, we report that culturing human pancreatic islets in the presence of serum resulted in the emergence of a population of nestin-positive cells. These proliferating cells were mainly C-peptide negative, although in the first week in culture, proliferating cells, insulin promoter factor-1 (Ipf-1) positive, were observed. Later passages of islet-derived cells were Ipf-1 negative and displayed a mesenchymal phenotype. These human pancreatic islet-derived mesenchymal (hPIDM) cells were expanded up to 1014 cells and were able to differentiate toward adipocytes, osteocytes and chondrocytes, similarly to mesenchymal stem/precursor cells. Interestingly, however, under serum-free conditions, hPIDM cells lost the mesenchymal phenotype, formed islet-like clusters (ILCs) and were able to produce and secrete insulin. These data suggest that, although these cells are likely to result from preexisting mesenchymal cells rather than β-cells, hPIDM cells represent a valuable model for further developments toward future replacement therapy in diabetes.
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Abbreviations
- ABCG2:
-
ATP-binding cassette sub-family G member 2
- aFABP:
-
adipocyte fatty acid binding protein
- AP:
-
alkaline phosphatase
- BrdU:
-
5-bromo-2′deoxyuridine
- BSA:
-
bovine serum albumin
- CBFA-1:
-
core binding factor alpha 1
- FACS:
-
fluorescence-activated cell sorting
- FBS:
-
fetal bovine serum
- GAPDH:
-
glyceraldehyde-3-phosphate dehydrogenase
- GFP:
-
green fluorescent protein
- GLUT-2:
-
glucose transporter-2
- GM:
-
growth medium
- hPIDM:
-
human pancreatic islet-derived mesenchymal
- HPRT:
-
hypoxanthine guanine phosphoribosyl transferase
- hTERT:
-
human telomerase reverse transcriptase
- ILCs:
-
islet-like clusters
- Ipf-1:
-
insulin promoter factor-1
- ISL-1:
-
islet-1
- NEUROD1:
-
neurogenic differentiation 1
- NKX2-2:
-
NK2 transcription factor-related locus2
- NKX6-1:
-
NK6 transcription factor related locus1
- PC1, 3:
-
prohormone convertase 1, 3
- PPARγ2:
-
peroxisome proliferator-activated receptor γ 2
- α-SMA:
-
α-smooth muscle actin
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Acknowledgements
This work was supported by a core grant from Regione Toscana to the Center for Stem Cell Research, University of Siena, and by grants from the Italian Ministry of Health and from the Umberto Di Mario ONLUS Foundation.
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Gallo, R., Gambelli, F., Gava, B. et al. Generation and expansion of multipotent mesenchymal progenitor cells from cultured human pancreatic islets. Cell Death Differ 14, 1860–1871 (2007). https://doi.org/10.1038/sj.cdd.4402199
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DOI: https://doi.org/10.1038/sj.cdd.4402199
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