2. Academic Validation
  3. Harnessing reaction-based probes to preferentially target pancreatic β-cells and β-like cells

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

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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