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Effective Islet Isolation Method With Extremely High Islet Yields From Adult Pigs

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Abstract:

Achieving good islet isolation is one of the most important factors for successful islet transplantation. Porcine pancreas is suitable for islet isolation research due to its anatomical and physiological similarities to human pancreas. In this study, we evaluated a new porcine islet isolation method designed to maximize islet yield and compared it with our previous open pan method and the standard method using a Ricordi chamber (Ricordi method). We performed 15 porcine islet isolations, five each with the new method, the open pan method, and the Ricordi method. The new method features several important improvements. Pancreata remain uncut and are kept intact during collagenase intraductal injection, a large filtration chamber to handle whole pancreata, low concentration of collagenase (LiberaseTM HI) for digestion, and large plastic containers for large-scale islet purification. All isolated islets were assessed for yield, purity, viability and in vitro function. Islets isolated with this new method were transplanted under the kidney capsules of SCID mice with chemically induced diabetes for in vivo functional assessment (n = 8). With the new method, we obtained on average more than 1,000,000 islet equivalents (IE) (1,236,266 ± 213,486 IE) (mean ± SE) before purification and 800,000 IE (879,815 ± 222,729 IE) after purification from one adult pig. Islet yield per pancreas was significantly higher compared with our previous open pan method (30,666 ± 11,532 IE, p < 0.01) and the Ricordi method (317,073 ± 86,093 IE, p < 0.05). All mice, transplanted with 1000 islets from the new method, returned to normoglycemia within 4 days after transplantation. Our new method makes it possible to obtain extremely high porcine islet yield with good function. It should produce useful information for human islet isolation and transplantation, and might be applied to single donor clinical xenogeneic transplantation.

Keywords: Intraductal injection; Islet transplantation; Porcine islets; Whole pancreas; Xenotransplantation

Document Type: Research Article

DOI: http://dx.doi.org/10.3727/000000005783982512

Affiliations: 1: Department of Transplantation and Immunology, Graduate School of Medicine, Kyoto University, 54 Kawara-cho Shogoin, Sakyo-ku, Kyoto, 606-8507 Japan 2: Kyoto University Hospital, Transplantation Unit, 54 Kawara-cho Shogoin, Sakyo-ku, Kyoto, 606-8507 Japan 3: Johnson & Johnson, 199 Grandview Road, Skillman, NJ 08558, USA 4: Department of Transplantation and Immunology, Graduate School of Medicine, Kyoto University, 54 Kawara-cho Shogoin, Sakyo-ku, Kyoto, 606-8507 Japan, Kyoto University Hospital, Transplantation Unit, 54 Kawara-cho Shogoin, Sakyo-ku, Kyoto, 606-8507 Japan

Publication date: October 1, 2005

More about this publication?
  • Cell Transplantation publishes original, peer-reviewed research and review articles on the subject of cell transplantation and its application to human diseases. To ensure high-quality contributions from all areas of transplantation, separate section editors and editorial boards have been established. Articles deal with a wide range of topics including physiological, medical, preclinical, tissue engineering, and device-oriented aspects of transplantation of nervous system, endocrine, growth factor-secreting, bone marrow, epithelial, endothelial, and genetically engineered cells, among others. Basic clinical studies and immunological research papers are also featured. To provide complete coverage of this revolutionary field, Cell Transplantation will report on relevant technological advances, and ethical and regulatory considerations of cell transplants. Cell Transplantation is now an Open Access journal starting with volume 18 in 2009, and therefore there will be an inexpensive publication charge, which is dependent on the number of pages, in addition to the charge for color figures. This will allow work to be disseminated to a wider audience and also entitle the corresponding author to a free PDF, as well as prepublication of an unedited version of the manuscript.
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