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A human β-cell line for transplantation therapy to control type 1 diabetes

Nature Biotechnology volume 23, pages 1274–1282 (2005)Cite this article

Abstract

A human pancreatic β-cell line that is functionally equivalent to primary β-cells has not been available. We established a reversibly immortalized human β-cell clone (NAKT-15) by transfection of primary human β-cells with a retroviral vector containing simian virus 40 large T-antigen (SV40T) and human telomerase reverse transcriptase (hTERT) cDNAs flanked by paired loxP recombination targets, which allow deletion of SV40T and TERT by Cre recombinase. Reverted NAKT-15 cells expressed β-cell transcription factors (Isl-1, Pax 6, Nkx 6.1, Pdx-1), prohormone convertases 1/3 and 2, and secretory granule proteins, and secreted insulin in response to glucose, similar to normal human islets. Transplantation of NAKT-15 cells into streptozotocin-induced diabetic severe combined immunodeficiency mice resulted in perfect control of blood glucose within 2 weeks; mice remained normoglycemic for longer than 30 weeks. The establishment of this cell line is one step toward a potential cure of diabetes by transplantation.

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Figure 1: Scheme for the establishment of reversibly immortalized human β-cell lines.
Figure 2: Molecular characterization of a human pancreatic β-cell line, NAKT-15.
Figure 3: Morphological and immunohisto-chemical analysis of reverted NAKT-15 cells.
Figure 4: Remission of diabetes by transplantation of reverted NAKT-15 cells into diabetic SCID mice.
Figure 5: Characterization of the reverted NAKT-15 cells after transplantation into diabetic mice.
Figure 6: Histological analysis of the kidney capsule, pancreas and liver of SCID mice transplanted with NAKT-15 cells.

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Acknowledgements

The work presented in this paper was supported in part by a Grant-in-Aid for Scientific Research (B) of the Japan Society for the Promotion of Science to N.K., Life Science Project of 21st Century, Japan, to N.T., and the American Diabetes Association (1-04-ISLET-31) and National Institutes of Health grant 1R21DK60192 to J.W.Y. and H.S.J. We thank Ann Kyle for editorial assistance.

Author information

Authors and Affiliations

  1. Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan

    Michiki Narushima, Naoya Kobayashi, Yong Chen, Atsushi Miki, Kimiaki Tanaka, Alejandro Soto Gutierrez, Jorge David Rivas-Carrillo, Nalu Navarro-Álvarez & Noriaki Tanaka

  2. Department of Transplant Surgery, Kyoto University Hospital, 54 Seigoin-Kawaracho, Sakyoku, Kyoto, 606-8507, Japan

    Teru Okitsu & Hirofumi Noguchi

  3. Medical Products Department, Kuraray Medical Co. Ltd., 1621 Shakazu, Kurashiki, 710-8622, Japan

    Yoshihito Tanaka & Shuhei Nakaji

  4. Department of Neuroscience, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama, 700-8558, Japan

    Shun-Ai Li & Kohji Takei

  5. Rosalind Franklin Comprehensive Diabetes Center, Chicago Medical School, 3333 Green Bay Road, North Chicago, 60064, Illinois, USA

    Hee-Sook Jun & Ji-Won Yoon

  6. Department of Biochemistry, Chosun University School of Medicine, 375 Seosuk-Dong, Gwangju, 501-759, Korea

    Hee-Sook Jun

  7. Division of Health Sciences and Technology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Karen A Westerman & Philippe Leboulch

  8. Genetix Pharmaceuticals, 840 Memorial Drive, Cambridge, 02139, Massachusetts, USA

    Karen A Westerman & Philippe Leboulch

  9. Human Pancreatic Islet Transplant Program, The University of Alberta, T2N 4N1, Alberta, Canada

    Jonathan R T Lakey

  10. Harvard Medical School and Division of Hematology, Department of Medicine, Brigham and Women's Hospital, 75 Francis Street, Boston, 02115, Massachusetts, USA

    Philippe Leboulch

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  1. Michiki Narushima
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Correspondence to Naoya Kobayashi or Ji-Won Yoon.

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Supplementary information

Supplementary Fig. 1

Quantification of northern and western blots of islet-specific genes in immortalized and reverted NAK-15 cells. (PDF 109 kb)

Supplementary Fig. 2

Real-time PCR analysis of islet-specific genes in immortalized and reverted NAKT-15 cells. (PDF 74 kb)

Supplementary Fig. 3

Loss of proliferation and lack of tumorigenicity of reverted NAKT-15 cells. (PDF 93 kb)

Supplementary Fig. 4

Insulin secretion and content of NAKT-15 cells reverted at different times during passage in culture. (PDF 64 kb)

Supplementary Table 1

Gene expression of β cell-specific transcription factors and insulin in 253 clones of immortalized human β cells. (PDF 27 kb)

Supplementary Table 2

Insulin secreted in response to glucose and insulin and C-peptide content in reverted NAKT-15 cells in vitro (PDF 67 kb)

Supplementary Table 3

Insulin secreted in response to various secretagogues in reverted NAKT-15 cells in vitro. (PDF 72 kb)

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Narushima, M., Kobayashi, N., Okitsu, T. et al. A human β-cell line for transplantation therapy to control type 1 diabetes. Nat Biotechnol 23, 1274–1282 (2005). https://doi.org/10.1038/nbt1145

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