A differentiation protocol for generating pancreatic delta cells from human pluripotent stem cells

Area of Science:

• Stem cell biology
• Endocrinology
• Cell differentiation

Background:

• Pancreatic delta cells play a crucial role in glucose homeostasis.
• Generating functional delta cells from human pluripotent stem cells (hPSCs) is essential for research and therapeutic applications.
• Existing differentiation protocols can be complex and inefficient.

Purpose of the Study:

• To detail a robust and simplified seven-stage protocol for generating pancreatic delta cells (SC-delta cells) from hPSCs.
• To provide a stable and efficient source of SC-delta cells for further investigation.
• To facilitate the refinement of islet cell therapeutic strategies.

Main Methods:

• Induction of definitive endoderm using activin A and CHIR99021.
• Sequential treatment with specific growth factors and small molecules (FGF7, SANT1, LDN193189, PdBU, retinoic acid, FGF2, XXI, ALK5 inhibitor II, Betacellulin) to guide differentiation through primitive gut tube, posterior foregut, and endocrine stages.
• Transition from planar to suspended culture after stage 5 to promote spheroid formation of delta cells.

Main Results:

• Successful generation of pancreatic delta cells (SC-delta cells) from hPSCs within approximately 4-5 weeks.
• Additional 1-2 weeks for cell expansion and evaluation.
• The protocol yields a stable source of SC-delta cells.

Conclusions:

• This seven-stage protocol offers an amenable and simplified method for efficient SC-delta cell differentiation.
• The generated SC-delta cells can advance the understanding of delta cell physiology.
• This protocol supports the development of improved islet cell therapeutic strategies.