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Maturation and function of human embryonic stem cell-derived pancreatic progenitors in macroencapsulation devices following transplant into mice

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Abstract

Aims/hypothesis

Islet transplantation is a promising cell therapy for patients with diabetes, but it is currently limited by the reliance upon cadaveric donor tissue. We previously demonstrated that human embryonic stem cell (hESC)-derived pancreatic progenitor cells matured under the kidney capsule in a mouse model of diabetes into glucose-responsive insulin-secreting cells capable of reversing diabetes. However, the formation of cells resembling bone and cartilage was a major limitation of that study. Therefore, we developed an improved differentiation protocol that aimed to prevent the formation of off-target mesoderm tissue following transplantation. We also examined how variation within the complex host environment influenced the development of pancreatic progenitors in vivo.

Methods

The hESCs were differentiated for 14 days into pancreatic progenitor cells and transplanted either under the kidney capsule or within Theracyte (TheraCyte, Laguna Hills, CA, USA) devices into diabetic mice.

Results

Our revised differentiation protocol successfully eliminated the formation of non-endodermal cell populations in 99% of transplanted mice and generated grafts containing >80% endocrine cells. Progenitor cells developed efficiently into pancreatic endocrine tissue within macroencapsulation devices, despite lacking direct contact with the host environment, and reversed diabetes within 3 months. The preparation of cell aggregates pre-transplant was critical for the formation of insulin-producing cells in vivo and endocrine cell development was accelerated within a diabetic host environment compared with healthy mice. Neither insulin nor exendin-4 therapy post-transplant affected the maturation of macroencapsulated cells.

Conclusions/interpretation

Efficient differentiation of hESC-derived pancreatic endocrine cells can occur in a macroencapsulation device, yielding glucose-responsive insulin-producing cells capable of reversing diabetes.

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Abbreviations

CXCR4:

Chemokine [C-X-C motif] receptor 4

GLP-1R:

Glucagon-like peptide 1 receptor

H&E:

Haematoxylin and eosin

hESCs:

Human embryonic stem cells

MAFA:

v-Maf musculoaponeurotic fibrosarcoma oncogene homolog A (avian)

NKX2.2:

NK2 homeobox 2

NKX6.1:

NK6 homeobox 1

S4:

Stage 4

STZ:

Streptozotocin

UBC:

University of British Columbia

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Acknowledgements

This work was funded by the Canadian Institutes of Health Research (CIHR) Regenerative Medicine and Nanomedicine Initiative, Stem Cell Network (SCN), JDRF, and Stem Cell Technologies. We would like to thank Ali Asadi for his technical assistance with immunofluorescent staining.

Funding

TJK was supported by a senior scholarship from the Michael Smith Foundation for Health Research and received financial support from Janssen R&D LLC. JEB was funded by a JDRF postdoctoral fellowship, a CIHR postdoctoral fellowship and the CIHR Transplantation Training Program. JEB also received a L’Oréal Canada for Women in Science Research Excellence Fellowship.

Duality of interest

AR, JX, KN and JJO’N are employees of Janssen R&D LLC, and TJK received financial support from Janssen R&D LLC for the research described in this article. The other authors confirm that there is no duality of interest associated with their contribution to this manuscript.

Contribution statement

JEB wrote the manuscript. AR, JEB, JX and TJK contributed to conception and design of experiments. AR, JEB, JX, KN, JJO’N and JKF were responsible for the acquisition, analysis and interpretation of data. All authors contributed to manuscript revisions and approved the final version of the manuscript.

Author information

Authors and Affiliations

  1. Laboratory of Molecular and Cellular Medicine, Department of Cellular & Physiological Sciences, Life Sciences Institute, University of British Columbia, Room 5308-2350 Health Sciences Mall, Vancouver, BC, Canada, V6T 1Z3

    Jennifer E. Bruin, Jessica K. Fox & Timothy J. Kieffer

  2. BetaLogics Venture, Janssen R&D LLC, Raritan, NJ, USA

    Alireza Rezania, Jean Xu, Kavitha Narayan & John J. O’Neil

  3. Department of Surgery, University of British Columbia, Vancouver, BC, Canada

    Timothy J. Kieffer

Authors
  1. Jennifer E. Bruin
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  2. Alireza Rezania
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  3. Jean Xu
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  4. Kavitha Narayan
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  5. Jessica K. Fox
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  6. John J. O’Neil
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  7. Timothy J. Kieffer
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Corresponding author

Correspondence to Timothy J. Kieffer.

Additional information

Jennifer E. Bruin and Alireza Rezania contributed equally to this work.

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Bruin, J.E., Rezania, A., Xu, J. et al. Maturation and function of human embryonic stem cell-derived pancreatic progenitors in macroencapsulation devices following transplant into mice. Diabetologia 56, 1987–1998 (2013). https://doi.org/10.1007/s00125-013-2955-4

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