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Hong, Han, Kim, Park, Lee, Jeong, Min, Lee, Kim, and Lee: Activin A and Glucose Derived Human Pancreatic Ductal Cells into Insulin-producing Cells
Original Article
The Journal of Korean Diabetes Association

The Journal of Korean Diabetes Association 2007; 31(1): 44-50.

Published online: 31 January 2007

DOI: https://doi.org/10.4093/jkda.2007.31.1.44

Activin A and Glucose Derived Human Pancreatic Ductal Cells into Insulin-producing Cells

Seung Hyun Hong1, 2, Chul Han1, 3, Hyo Sup Kim1, Mi Kyung Park1, Young Jin Lee1, Jae Hoon Jeong1, 3, Yong Ki Min1, 3, Myung Shik Lee1, 3, Kwang Won Kim1, 3, Moon Kyu Lee1, 3

1Samsung Biomedical Research Institute (SBRI), Korea.

2Department of Life Science, College of Natural Sciences, Chungang University, Korea.

3Division of Endocrinology and Metabolism, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Korea.

Received 12 September 2006       Accepted 25 January 2007

Copyright © 2007 Korean Diabetes Association

Abstract

Background

Cellular replacement therapy holds promise for the treatment of diabetes mellitus but donor tissue is severely limited. Human postnatal pancreatic ductal cells are a potential source of new beta cells. Therefore, we investigated the potential of human pancreatic ductal cells could be differentiated into endocrine cells that would be capable of secreting insulin in response to glucose.

Methods

Cell fractions enriched with pancreatic ductal cells after human islet isolation were treated with streptozotocin to remove residual beta cells, grown in monolayer culture, changed the media for differentiation in the presence of activin A and glucose, supplemented with 10% FCS. The differentiation markers, insulin secretion and cell proliferation were examined.

Result

No insulin was detectable in cell preparations after 5 days of treatment with streptozotocin. In monolayer culture, 80% of the streptozotocin-treated pancreatic ductal cells expressed cytokeratin-19. Cell cultures with a high proportion of cytokeratin-19 cells had greater plasticity for differentiation into cells with phenotypic and functional markers of beta cells. This property were significantly enhanced by treatment of activin A and glucose. The differentiated human pancreatic ductal cells secreted insulin sensitively responded with high glucose.

Conclusion

Human pancreatic ductal cells are a potential source of new glucose-induced insulin producing cells that may be developed further for clinical use. Therefore, the present data support a possible role for human adult pancreatic ductal cells, following expansion and differentiation, as a source of insulin by transplantation cells to type I diabetes patients.

Keywords: Activin A, Ductal cell

Figures and Tables

Fig. 1
Removal of beta cell from pancreatic ductal cell. A, Reduction of insulin mRNA during STZ treatment (n = 4). Variation of insulin mRNA expression of STZ-treated ductal cell compaired with STZ-nontreated ductal cell; B, RT-PCR showing preproinsulin and CK19 mRNA expression in the cells following STZ treatment; C, Cell constitution after STZ treatment.
*P < 0.05 vs. STZ nontreated ducatal cell.
jkda-31-44-g001
Fig. 2
Morphological charateristic of pancreatic ductal cell.
A, Complete Monolayer after STZ treatment for 5 days; B, Cluster formation after activin A and glucose treatment for 30 days.
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Fig. 3
Effect of activin A on the differentiation of ductal cell.
A, The mRNA expression of differentiation marker of activin A and glucose treated or nontreated ductal cells for 30 days; B, To characterize the cells of dispersed duct cell, triple immuno-staining with CK19, inslulin and DAPI was performed. Duct cells stained with CK19 (red). Beta cells stained with insulin (green) b, STZ treated ductal cell for 5 days. c, Activin A and glucose treated ductal cell for 15 days after STZ treatment. d, Activin A and glucose treated ductal cell for 30 days after STZ treatment; C, The mRNA expression of insulin and preproinsluin during differentiation.
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Fig. 4
Variation of glucose-induced insulin secretion during differentiation.
*P < 0.05 vs. glucose 3 mM.
jkda-31-44-g004
Fig. 5
Variation of cell growth during differentiation.
A, Doubling times were calculated using the Schwartz equation; B, The proliferation rates were assayed by the rate of BrdU incorporation.
jkda-31-44-g005
Table 1
Sequences of Primers Used in RT-PCR
jkda-31-44-i001

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