Differentiation of Affinity-Purified Human Pancreatic Duct Cells to β-Cells

  1. Shigeru Yatoh,
  2. Rikke Dodge,
  3. Tomoyuki Akashi,
  4. Abdulkadir Omer,
  5. Arun Sharma,
  6. Gordon C. Weir and
  7. Susan Bonner-Weir
  1. From the Section on Islet Transplantation and Cell Biology, Joslin Diabetes Center, Boston, Massachusetts
  1. Address correspondence and reprint requests to Susan Bonner-Weir, PhD, Section on Islet Transplantation and Cell Biology, Joslin Diabetes Center, One Joslin Place, Boston, MA 02215. E-mail: susan.bonner-weir{at}joslin.harvard.edu

Abstract

To test whether pancreatic duct cells are in vitro progenitors, they were purified from dispersed islet-depleted human pancreatic tissue using CA19-9 antibody. The purified fraction was almost entirely CK19+ with no insulin+ cells, whereas the unpurified cells (crude duct) were 56% CK19+ and 0.4% insulin+ of total cells (0.7% of CK19+ cells). These cells were expanded as monolayers, aggregated under serum-free conditions, and transplanted into normoglycemic NOD/SCID mice. In crude duct grafts, insulin+ cells increased to 6.1% of CK19+ cells. Purified duct cells had slow expansion and poor aggregation, as well as engraftment. The addition of 0.1% cultured stromal cells improved these parameters. These stromal cells contained no CK19+ cells and no insulin by either quantitative RT-PCR or immunohistochemistry; stromal cell aggregates and grafts contained no insulin+ cells. Aggregation of purified duct plus stromal preparations induced insulin+ cells (0.1% of CK19+ cells), with further increase to 1.1% in grafts. Insulin mRNA mirrored these changes. In these grafts, all insulin+ cells were in duct-like structures, while in crude duct grafts, 85% were. Some insulin+ cells coexpressed duct markers (CK19 and CA19-9) and heat shock protein (HSP)27, a marker of nonislet cells, suggesting the transition from duct. Thus, purified duct cells from adult human pancreas can differentiate to insulin-producing cells.

Footnotes

  • Published ahead of print at http://diabetes.diabetesjournals.org on 1 May 2007. DOI: 10.2337/db06-1670.

  • S.Y. is currently affiliated with the Department of Medicine (Metabolism and Endocrinology), University of Tsukuba, Japan.

  • The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

    • Accepted April 14, 2007.
    • Received November 29, 2006.
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  1. Diabetes vol. 56 no. 7 1802-1809
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