Pancreatic WT

DefiniGEN's wild-type pancreatic beta cells are functional human pancreatic cells displaying many of the characteristics of primary human beta cells. The resulting cells display a robust Glucose Sensitive Insulin Secretion (GSIS) response over a range of physiologically relevant glucose concentrations and exhibit an elevated GSIS response to key reference drugs in a similar manner to primary human pancreatic islets. Def-PANC WT cells can be grown in monolayer, or they can be conveniently cultured as microislets which resemble primary human pancreati islets in structure and function. The cells are available for key applications including drug discovery and Diabetes research.

• Highly standardized cell product containing >97% human pancreatic cells with consistent performance and biologically relevant data
• Wild-type donor genetics and karyotype verified
• Glucose sensitive insulin secretion (GSIS) response over a range of physiologically relevant glucose concentrations
• Dose dependent GSIS response to key diabetes reference drugs

Quantification of the pancreatic insulin gene expression marker by qPCR

Def-PANC cells express the insulin gene at very similar levels to primary human pancreatic islets.

Figure 1.Insulin gene expression marker analysis. Insulin QPCR-analysis observed in Def-PANC cells thawed from a cryopreserved vial in 96 well low adherent plates and cultured as microislets relative to control primary islets.

Glucose Stimulated Insulin Secretion Assay

A robust GSIS response is observed in Def-PANC cells when thawed from a cryopreserved vial in 96 well low adherent plates and grown as islet-like structures. They have also demonstrated a dose-dependent response to well-known secretagogues such as GLP-1 and Exenatide.

Figure 2. Def-PANC WT cells display a robust GSIS response. Panel (A) shows GSIS assay results for Def-PANC cells stimulated with low and high concentrations of glucose only. Panel (B) overviews the GSIS response of Def-PANC cells stimulated with low and high concentrations of glucose and exenatide. Low glucose concentration 1.6mM, high glucose concentration 16.7mM, exenatide concentration 25nM.

Monolayer culture

The Def-PANC cell products are highly functional iPSC-derived pancreataic cells. Yamanaka iPSC technology in combination with fully defined differentiation conditions enables the generation of standardized populations of pancreatic cell products. Through a 25 day differentiation process the cells proceed through key developmental stages ultimately producing functional pancreatic cells (Figure 3).

A)                                                                          B)                                                                               C)

Figure 3. Pancreatic cell type distribution in monolayer. When grown in standard laboratory 96-well plates Def-PANC cells show typically tightly packed pancreatic cell morphology (A) and a high proportion of C-peptide secretion from beta cells (green) (B). Panel C depicts DAPI staining on nuclear DNA (blue) as well as C-peptide (green).

Microislet culture

When grown on 96-well low adherent plates the Def-PANC cells aggregate and form microislet structures of similar size to primary human islets (Figure 4).

Figure 4. Immunostaining analysis of Def-PANC cells. Microislet formation of Def-PANC in low adherent plates. Insulin secreting beta cells (green) are the dominant cell population observed alongside lower populations of glucagon and somatostatin-expressing cells (size 100-150 um).