A Method for Organoid Transplantation and Whole-Mount Visualization of Post-Engraftment Vascularization

Methods Mol Biol. 2021:2258:259-272. doi: 10.1007/978-1-0716-1174-6_17.

Abstract

As the field of organoid development matures, the need to transplant organoids to evaluate and characterize their functionality grows. Decades of research developing islet organoid transplantation for the treatment of type 1 diabetes can contribute substantially to accelerating diverse tissue organoid transplantation. Biomaterials-based organoid delivery methods offer the potential to maximize organoid survival and engraftment. In this protocol, we describe a vasculogenic degradable hydrogel vehicle and a method to deliver organoids to intraperitoneal tissue. Further, we describe a method to fluorescently label and image functional vasculature within the graft as a tool to investigate organoid engraftment.

Keywords: Engraftment; Organoids; Transplantation; Vascularization.

MeSH terms

  • Angiogenesis Inducing Agents / pharmacology
  • Animals
  • Cell Culture Techniques
  • Cells, Cultured
  • Genes, Reporter
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Hydrogels
  • Islets of Langerhans / blood supply*
  • Islets of Langerhans / metabolism
  • Islets of Langerhans Transplantation
  • Maleimides / chemistry
  • Mice
  • Microscopy, Confocal*
  • Neovascularization, Physiologic* / drug effects
  • Organoids / blood supply*
  • Organoids / metabolism
  • Organoids / transplantation*
  • Polyethylene Glycols / chemistry
  • Rats
  • Tissue Engineering*
  • Tissue Fixation
  • Vascular Endothelial Growth Factor A / pharmacology

Substances

  • Angiogenesis Inducing Agents
  • Hydrogels
  • Maleimides
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Green Fluorescent Proteins
  • Polyethylene Glycols