We exploited the anterior chamber of the eye (ACE) of immunodeficient mice as an ectopic site for both transplantation and microimaging of engineered surrogate islets from human induced pluripotent stem cells (hiPSC-SIs). These islets contained a majority of insulin-expressing cells, positive or negative for PDX1 and NKX6.1, and a minority of glucagon- or somatostatin-positive cells. Single, non-aggregated hiPSC-SIs were satisfactorily engrafted onto the iris. They underwent gradual vascularization and progressively increased their light scattering signals, reflecting the abundance of zinc-insulin crystal packaged inside mature insulin secretory granules. Intracameral hiPSC-SIs retrieved from recipients showed enhanced insulin immunofluorescence in correlation with the parallel increase in overall vascularization and light backscattering during the post-transplantation period. This approach enables longitudinal, nondestructive and intravital microimaging of cell fates, engraftment, vascularization and mature insulin secretory granules of single hiPSC-SI grafts, and may offer a feasible and reliable means to screen compounds for promoting in vivo hiPSC-SI maturation
Bibliographical noteFunding Information:
P-OB is the founder and CEO of the biotech company BioCrine AB. S-NY is a consultant to BioCrine. AK and JO are co-founders of Regenerative Medical Solutions, Inc. and have equity interest in the company. The research reported here was supported in part by funding provided by Regenerative Medical Solutions, Inc. AM, and TJ are employees of Regenerative Medical Solutions, Inc.
The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This research was supported by The Bert von Kantzow Foundation, Diabetes and Wellness Foundation, The ERC-2013-AdG 338936-BetaImage, The Family Erling-Persson Foundation, The Family Knut and Alice Wallenberg Foundation, KI Foundations & Funds, Skandia Insurance Company, Ltd., Strategic Research Program in Diabetes at Karolinska Institutet, The Swedish Diabetes Association, The Swedish Research Council and The Stichting af Jochnick Foundation. RMS has been supported by NIH (NIDDK) SBIR grants (2R44DK104497-02, 1R43DK104497-01, 1R43DK108441-01A1, 1R43DK112472-01, 5R44DK104497-03 and 1R43DK109832-01A1).
© The Author(s) 2022.
- human induced pluripotent stem cells (hiPSCs)
- in vivo confocal microscopy
- surrogate islet
- the anterior chamber of the eye (ACE)