Maturation and function of human embryonic stem cell-derived pancreatic progenitors in macroencapsulation devices following transplant into mice
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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.
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.
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.
Efficient differentiation of hESC-derived pancreatic endocrine cells can occur in a macroencapsulation device, yielding glucose-responsive insulin-producing cells capable of reversing diabetes.
KeywordsCell therapy Diabetes Encapsulation Human embryonic stem cells Insulin Islets
Chemokine [C-X-C motif] receptor 4
Glucagon-like peptide 1 receptor
Haematoxylin and eosin
Human embryonic stem cells
v-Maf musculoaponeurotic fibrosarcoma oncogene homolog A (avian)
NK2 homeobox 2
NK6 homeobox 1
University of British Columbia
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.
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.
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.
- 53.Zhu X, Orci L, Carroll R, Norrbom C, Ravazzola M, Steiner DF (2002) Severe block in processing of proinsulin to insulin accompanied by elevation of des-64,65 proinsulin intermediates in islets of mice lacking prohormone convertase 1/3. Proc Natl Acad Sci U S A 99:10299–10304PubMedCrossRefGoogle Scholar