STEMdiff™ Pancreatic Progenitor Kit
Serum-free medium for the differentiation of human ES and iPS cells to pancreatic progenitor cells
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- Y-27632 (Dihydrochloride)
RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
- D-PBS (Without Ca++ and Mg++)
Dulbecco’s phosphate-buffered saline without calcium and magnesium
- Gentle Cell Dissociation Reagent
cGMP, enzyme-free cell dissociation reagent
STEMdiff™ Pancreatic Progenitor Kit has been optimized for differentiation of cells maintained in mTeSR™1 (Catalog #85850).
Figure 1. Pancreatic Progenitor Cells can Mature into Endocrine and Exocrine Cells
A) Representative image showing pancreatic progenitor cells expressing PDX-1 (red) and NKX6.1 (green). Yellow staining indicates co‑expression of both markers in the majority of cells as is observed in the developing human pancreas.1 (B, C) Cells transplanted into mice can mature into endocrine and exocrine cells.
B) shows endocrine clusters expressing synaptophysin (red) surrounded by ductal structures expressing CK-19 (green).
C) Shows islet-like structures containing monohormonal cells that individually express insulin (red), glucagon (green) or somatostatin (blue). Data in (B, C) are from the laboratory of Dr. Timothy J. Kieffer (University of British Columbia, Vancouver, Canada).
Figure 2. The STEMdiff™ Pancreatic Progenitor Kit Functions Efficiently Across Multiple hPSC Lines
PDX-1 and NKX6.1 expression measured in pancreatic progenitor cells derived from four different hPSC lines (H1, H9, WLS-4D1 and WLS-1C) at the end of Stage 4. (A) Representative flow cytometry plots show PDX-1 and NKX6.1 co-expression in differentiated H9 cells. (B) Quantitative data for PDX-1/NKX6.1 co-expression in two human ES (H1 and H9) and two human iPS (WLS-4D1 and WLS-1C) cell lines (n = 5-10 per cell line). Data are plotted as individual points representing the mean of duplicates within a single experiment. The horizontal line represents the mean of all experiments, with error bars indicating the standard error of the mean (SEM). The average efficiency of pancreatic progenitor differentiation ranges from 61.5% to 77.7% depending on the cell line.
Figure 3. Gene Expression Profile is Indicative of Transition to Pancreatic Progenitor Cells
Gene expression profile at the end of each stage of differentiation for key markers of pancreatic progenitor cells. Expression was normalized to 18S ribosomal RNA and TATA Binding Protein (TBP). Data are the mean ± SEM for 3 - 5 experiments. Expression pattern is consistent with published data.²
1. Riedel M et al. (2012) Immunohistochemical characterization of cells co-producing insulin and glucagon in the developing human pancreas. Diabetologia 55(2): 372-81.
2. Rezania A et al. (2014) Reversal of diabetes with insulin-producing cells derived in vitro from human pluripotent stem cells. Nat Biotechnol 32(11): 1121-33.
Figure 4. Generation of Pancreatic Progenitors from hPSCs Maintained in mTeSR™ Plus
(A) Representative density plots showing PDX-1 and NKX6.1 expression in cells cultured in mTeSR™1 (daily feeds) or mTeSR™ Plus (restricted feeds), following differentiation using the STEMdiff™ Pancreatic Progenitor Kit. (B) Quantitative analysis of pancreatic progenitor formation in multiple hPS (H9, STiPS-M001, WLS-1C) cell lines maintained with mTeSR™1 or mTeSR™ Plus as measured by co-expression of PDX-1 and NKX6.1. Data are expressed as the mean percentage of cells (± SEM) expressing both markers; n=3.
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (10)
We get a really great efficiency of differentiation in several pluripotent stem cell lines determined by expression of key markers: upregulation of PDX1, SOX9, FOXA2 and GATA4, and down-regulation of Sox17. We can generate these pancreatic progenitor cells reproducibly and efficiently; variability within the protocol is low.
Drs. Jamie Trott and Ray Dunn, A*STAR Institute of Medical Biology (IMB), Singapore
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STEMdiff™ Pancreatic Progenitor Kit
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.