Human pancreatic islet-derived extracellular vesicles modulate insulin expression in 3D-differentiating iPSC clusters

Area of Science:

• Cell Biology
• Stem Cell Research
• Endocrinology

Background:

• Extracellular vesicles (EVs) mediate intercellular communication in pancreatic tissue.
• The role of pancreatic EVs in stem cell differentiation into pancreatic lineages is not well understood.

Purpose of the Study:

• To investigate the effect of human islet-derived EVs (h-Islet-EVs) on the differentiation of human induced pluripotent stem cells (iPSCs) into pancreatic lineages.
• To determine if h-Islet-EVs can regulate insulin and C-peptide expression during iPSC differentiation.

Main Methods:

• Human islet-derived EVs were isolated and characterized for size and EV markers (CD63, CD81).
• Presence of pancreatic transcription factors and hormones within h-Islet-EVs was confirmed.
• h-Islet-EVs were added to iPSC clusters during pancreatic differentiation protocols, including those in 3D-collagen hydrogels.

Main Results:

• h-Islet-EVs were characterized with a mean size of 117±7 nm and positive expression of CD63 and CD81.
• Key pancreatic markers (NGN3, MAFA, PDX1 mRNA; C-peptide, glucagon proteins) were detected in h-Islet-EVs.
• Supplementation with h-Islet-EVs upregulated intracellular C-peptide levels in iPSC clusters in a concentration-dependent manner.
• While 3D culture enhanced pancreatic marker mRNA, h-Islet-EVs did not significantly alter intracellular insulin or C-peptide content in this context.

Conclusions:

• h-Islet-EVs play a role in regulating insulin expression in differentiating iPSC clusters.
• These findings have significant implications for pancreatic tissue engineering and the development of cell-based therapies for diabetes.