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KMID : 0354720070310030236
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Journal of Korean Diabetes Association
2007 Volume.31 No. 3 p.236 ~ p.242
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Differentiation of Pancreatic β Cells from Human Pancreatic Duct Cells Derived from a Partial Pancreas Tissue
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Song Ki-Ho
Kim Myung-Mi
Lee Min-Kyung
Ryu Ryul-Gyeong
Ko Seung-Hyun
Moon Sung-Dae
Ahn Yu-Bae
Yoon Kun-Ho
Cha Bong-Yun
Lee Kwang-Woo
Son Ho-Young
Kang Sung-Koo
Chin Hyung-Min
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Abstract
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Background: Despite a recent breakthrough in human islet transplantation for treating diabetes mellitus, the
limited availability of insulin-producing tissue is still a major obstacle. This has led to a search for alternative sources of transplantable insulin-producing cells including pancreatic duct cells. We aimed to establish in vitro culture of pancreatic duct cells from a partial pancreas tissue in human, which could be harnessed to differentiate into pancreatic β cells.
Methods: We isolated pancreatic duct cells from small pieces of pancreas tissue (1~3 g) derived from non-diabetic humans (n = 8) undergoing pancreatic surgery due to cancer. Pancreas tissue was finely minced after injection of collagenase P into the parenchyma. The mince was incubated in a shaking water bath at 37℃ for 25 min and passed through a 150 μm mesh. The released cells were recovered, washed, and plated in a dish containing CMRL culture medium with serum.
Results: Isolated pancreatic cells grew in monolayer and became confluent in 1~2 wks showing typical epithelial cobblestone morphology. Immunochemistry demonstrated that ~90% of the cultured cells were cytokeratin7-positive duct cells. To induce β cell differentiation, the cells were incubated in DMEM/F12 culture medium without serum. In addition, treatment with Matrigel overlay, exendin-4, cholera toxin or forskolin was done. Though β cell differentiation was found by immunostaining and RT-PCR, the differentiation efficiency was very low. Over-expression of neurogenin-3 by recombinant adenovirus did not increase β cell differentiation of the cultured duct cells significantly.
Conclusion: We established in vitro culture of pancreatic duct cells from a partial pancreas tissue in human,
which differentiate into pancreatic cells. However, a strategy to optimize β cell differentiation in this model is needed.
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KEYWORD
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Beta cell, Differentiation, Islet, Islet transplantation, Pancreatic duct cells
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