Harnessing reaction-based probes to preferentially target pancreatic β-cells and β-like cells

  • Life Sci Alliance. 2021 Jan 29;4(4):e202000840. doi: 10.26508/lsa.202000840.
Sevim Kahraman  1 ,  Debasish Manna  2  3  4 ,  Ercument Dirice  1 ,  Basudeb Maji  2  3  4 ,  Jonnell Small  2  5 ,  Bridget K Wagner  2 ,  Amit Choudhary  6  3  4  5 ,  Rohit N Kulkarni  7
Affiliations
  • 1. Islet Cell and Regenerative Biology, Joslin Diabetes Center, Department of Medicine, Brigham and Women's Hospital, Harvard Stem Cell Institute, Harvard Medical School, Boston, MA, USA.
  • 2. Chemical Biology and Therapeutics Science Program, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
  • 3. Department of Medicine, Harvard Medical School, Boston, MA, USA.
  • 4. Divisions of Renal Medicine and Engineering, Brigham and Women's Hospital, Boston, MA, USA.
  • 5. Chemical Biology Program, Harvard University, Cambridge, MA, USA.
  • 6. Chemical Biology and Therapeutics Science Program, Broad Institute of MIT and Harvard, Cambridge, MA, USA [email protected].
  • 7. Islet Cell and Regenerative Biology, Joslin Diabetes Center, Department of Medicine, Brigham and Women's Hospital, Harvard Stem Cell Institute, Harvard Medical School, Boston, MA, USA [email protected].
PMID: 33514654 DOI: 10.26508/lsa.202000840
Abstract

Highly sensitive approaches to target insulin-expressing cells would allow more effective imaging, sorting, and analysis of pancreatic β-cells. Here, we introduce the use of a reaction-based probe, diacetylated Zinpyr1 (DA-ZP1), to image pancreatic β-cells and β-like cells derived from human pluripotent stem cells. We harness the high intracellular zinc concentration of β-cells to induce a fluorescence signal in cells after administration of DA-ZP1. Given its specificity and rapid uptake by cells, we used DA-ZP1 to purify live stem cell-derived β-like cells as confirmed by immunostaining analysis. We tested the ability of DA-ZP1 to image transplanted human islet grafts and endogenous mouse pancreatic islets in vivo after its systemic administration into mice. Thus, DA-ZP1 enables purification of insulin-secreting β-like cells for downstream applications, such as functional studies, gene-expression, and cell-cell interaction analyses and can be used to label engrafted human islets and endogenous mouse islets in vivo.

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