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. 2006 Jul 28;12(28):4529-35.
doi: 10.3748/wjg.v12.i28.4529.

Characterization of pancreatic stem cells derived from adult human pancreas ducts by fluorescence activated cell sorting

Han-Tso Lin  1 , Shih-Hwa Chiou, Chung-Lan Kao, Yi-Ming Shyr, Chien-Jen Hsu, Yih-Wen Tarng, Larry L-T Ho, Ching-Fai Kwok, Hung-Hai Ku
Affiliations

Characterization of pancreatic stem cells derived from adult human pancreas ducts by fluorescence activated cell sorting

Han-Tso Lin et al. World J Gastroenterol. .

Abstract

Aim: To isolate putative pancreatic stem cells (PSCs) from human adult tissues of pancreas duct using serum-free, conditioned medium. The characterization of surface phenotype of these PSCs was analyzed by flow cytometry. The potential for pancreatic lineage and the capability of beta-cell differentiation in these PSCs were evaluated as well.

Methods: By using serum-free medium supplemented with essential growth factors, we attempted to isolate the putative PSCs which has been reported to express nestin and pdx-1. The Matrigel(TM) was employed to evaluate the differential capacity of isolated cells. Dithizone staining, insulin content/secretion measurement, and immunohistochemistry staining were used to monitor the differentiation. Fluorescence activated cell sorting (FACS) was used to detect the phenotypic markers of putative PSCs.

Results: A monolayer of spindle-like cells was cultivated. The putative PSCs expressed pdx-1 and nestin. They were also able to differentiate into insulin-, glucagon-, and somatostatin-positive cells. The spectrum of phenotypic markers in PSCs was investigated; a similarity was revealed when using human bone marrow-derived stem cells as the comparative experiment, such as CD29, CD44, CD49, CD50, CD51, CD62E, PDGFR-alpha, CD73 (SH2), CD81, CD105(SH3).

Conclusion: In this study, we successfully isolated PSCs from adult human pancreatic duct by using serum-free medium. These PSCs not only expressed nestin and pdx-1 but also exhibited markers attributable to mesenchymal stem cells. Although work is needed to elucidate the role of these cells, the application of these PSCs might be therapeutic strategies for diabetes mellitus.

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Figures

Figure 1
Figure 1
Putative PSC isolated from adult pancreas. A: Morphology of cultivated putative PSCs; B: Cells aggregate when saturated. bar = 100 μm.
Figure 2
Figure 2
Nestin and pdx-1 expressed in putative PSCs after 5 passages of cultivation. cDNA oriented from 4 individual donors (lane No. 1 to 4), 100 bp marker (M) and the plasmid cloned human nestin and pdx-1 gene with positive control (C) were shown.
Figure 3
Figure 3
The differentiation of putative PSCs (4 wk). A: Growth in Matrigel™; B: Dithizone stain; C-E: Immunohistochemistry staining by anti-insulin (C), glucagon (D) and somatostatin (E) immunoglobulins. bar = 100 μm.
Figure 4
Figure 4
Measurement of Insulin content and secretion in differentiated putative PSCs.
Figure 5
Figure 5
Cell doubling time of putative PSC.
See this image and copyright information in PMC

References

    1. Treutelaar MK, Skidmore JM, Dias-Leme CL, Hara M, Zhang L, Simeone D, Martin DM, Burant CF. Nestin-lineage cells contribute to the microvasculature but not endocrine cells of the islet. Diabetes. 2003;52:2503–2512. - PubMed
    1. Shapiro AM, Lakey JR, Ryan EA, Korbutt GS, Toth E, Warnock GL, Kneteman NM, Rajotte RV. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen. N Engl J Med. 2000;343:230–238. - PubMed
    1. Humphrey RK, Bucay N, Beattie GM, Lopez A, Messam CA, Cirulli V, Hayek A. Characterization and isolation of promoter-defined nestin-positive cells from the human fetal pancreas. Diabetes. 2003;52:2519–2525. - PubMed
    1. Gao R, Ustinov J, Pulkkinen MA, Lundin K, Korsgren O, Otonkoski T. Characterization of endocrine progenitor cells and critical factors for their differentiation in human adult pancreatic cell culture. Diabetes. 2003;52:2007–2015. - PubMed
    1. Bonner-Weir S, Taneja M, Weir GC, Tatarkiewicz K, Song KH, Sharma A, O'Neil JJ. In vitro cultivation of human islets from expanded ductal tissue. Proc Natl Acad Sci USA. 2000;97:7999–8004. - PMC - PubMed

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