Enteroendocrine cell

Gene

Induced pluripotent stem cell

Actin

Internal medicine

Endocrinology

Cytoskeleton

Embryonic stem cell

Cellular differentiation

Cell

Islet

Insulin

Stem cell

Transplantation

Actin cytoskeleton

Endocrine system

Biochemistry

Hormone

Medicine

Biology

Cell biology

Nature Biotechnology

Generation of pancreatic β cells from human pluripotent stem cells (hPSCs) holds promise as a cell replacement therapy for diabetes. In this study, we establish a link between the state of the actin cytoskeleton and the expression of pancreatic transcription factors that drive pancreatic lineage specification. Bulk and single-cell RNA sequencing demonstrated that different degrees of actin polymerization biased cells toward various endodermal lineages and that conditions favoring a polymerized cytoskeleton strongly inhibited neurogenin 3-induced endocrine differentiation. Using latrunculin A to depolymerize the cytoskeleton during endocrine induction, we developed a two-dimensional differentiation protocol for generating human pluripotent stem-cell-derived β (SC-β) cells with improved in vitro and in vivo function. SC-β cells differentiated from four hPSC lines exhibited first- and second-phase dynamic glucose-stimulated insulin secretion. Transplantation of islet-sized aggregates of these cells rapidly reversed severe preexisting diabetes in mice at a rate close to that of human islets and maintained normoglycemia for at least 9 months.

Targeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells