Using Zinc-based probes to generate pseudoislets from purified human alpha-cells for functional analyses

Investigation of the function of human alpha-cells remains a challenge due to the lack of simple and cost-effective purification methods to isolate high-quality alpha-cells from islets. Here, we introduce a novel and simple method for enriching live alpha-cells from dissociated human islet cells by using the reaction-based probe diacetylated Zinpyr1 (DA-ZP1) with > 97% purity. Sorted islet cells from four islet donors were seeded in U-bottom low attachment 96 well plates (1000 cells/200 ul/well). Half of the medium was refreshed every other day for 10 days.