Generating functional pancreatic beta cells

Beta cell differentiation

Developments in diabetes research have advanced rapidly in recent years, including new cellular technologies for directed differentiation of human pluripotent stem cells (hPSCs) into pancreatic beta cells, with the goal of using these cells for cell replacement therapy to cure type I diabetes.

This protocol spotlight highlights the work of researchers from The Salk Institute who developed a novel strategy for directly differentiating pancreatic beta cells from hPSCs that is much more efficient than previous methods. The protocol involves the combination of chemical compounds and step-wise media supplementation to generate 3D pancreatic progenitor clusters from hPSCs, and further differentiation to NKX6.1+/INS+ beta cells. Researchers found that this protocol overcomes previous limitations for generating beta cells from hPSCs, with approximately 60% to 80% efficiency from multiple hPSC lines and origins. The publication further demonstrated the functionality of the beta cells generated using this protocol, showing that cells could rapidly reverse hyperglycemia in a mouse model of diabetes.

Read more about this innovative Nature Communications publication and corresponding differentiation protocol or review our specialized reagent list below to see how Captivate Bio can support diabetes research.

Figure: HPSC-derived PPs are dissociated and re-aggregated into PP 3D clusters by centrifuge and incubation in a V-shaped-bottom microplate and then transferred to the air–liquid interface. The PPs in 3D clusters mainly differentiate into β-cells after they are successively treated by PP-10C, EP-8C, iβ-9C, and Fβ-7C. However, if other suboptimal conditions are employed, the yield of β-cells is significantly reduced, whereas that of the dual-hormonal GCG+/INS+ cells increase. Schematic from data published in Liu, H., et al. Nat Commun. 2021.

Reagents

Cell Culture Products	Cat. No.	Size
DMEM/F12 Medium	DF1201	500 mL
MCDB 131 Medium	MCDB01	50 mL
RPMI 1640 Medium	RPMN01	500 mL
PBS 1X	PBS102	1000 mL
MEM-NEAA	NEAA03	100 mL
Accutase® Solution	CCAT03	100 mL
Compounds
CHIR99021	SML01B	10 mg
Compound E	SML02B	10 mg
Forskolin	SML19B	10 mg
LDN193189	SML05B	10 mg
RepSox	SML06B	10 mg
Retinoic Acid	SML07F	500 mg
SANT-1	SML08B	10 mg
Triiodothyronine Salt	SML11F	500 mg
Y-27632	SML13B	10 mg
H-1152	Please Inquire	10 mg
Nicotinamide (NICO)	Please Inquire	10 mg
L-Ascorbic Acid	Please Inquire	10 mg
PD173074	Please Inquire	10 mg
ISX-9	Please Inquire	10 mg
G-1	Please Inquire	10 mg
ZM447439	Please Inquire	10 mg
Supplementary Products
Human Platelet Lysate	View Portfolio	27 mL to 1000 mL
FBS, USDA Origin	FBS1620	500 mL
CET Cocktail	CET01B	1 Kit

References

• Liu, H., Li, R., Liao, HK. et al. Differentiation protocol for generating functional pancreatic β cells from human pluripotent stem cells, 08 June 2021, PROTOCOL (Version 1) available at Protocol Exchange
• Liu, H., Li, R., Liao, HK. et al. Chemical combinations potentiate human pluripotent stem cell-derived 3D pancreatic progenitor clusters toward functional β cells. Nat Commun 12, 3330 (2021).
• Research advances one step closer to stem cell therapy for type 1 diabetes – The Salk Institute

Captivate Bio Research and Protocol Spotlights are highlights written to summarize the utility of a study as well as showcasing the biological activity of a product. Always refer to the Authors and full publication or poster for more information.

Image source: This image shows functional beta cells made from human pluripotent stem cells. Insulin (red) and NKX6.1 (green) indicate two proteins produced by beta cells. The Salk Institute, June 2021.