Islets were isolated from mouse pancreas using perfusion with the Liberase enzyme blend (Sigma Aldrich), as described (21 (link)). For cell sorting, pancreatic buds from Pdx1-Cre:Rosa26R-YFP embryos, heterozygous Ngn3-EGFP embryos, or the MIP-GFP transgenic pups were harvested and digested into single cells using TrypLE Express (ThermoFisher). Cells were sorted for green fluorescent protein imaging using a BD FACS Aria II to an average percentage purity of 85–95%, with wild-type cells being the negative control for FACS gating (9 (link)).
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Isolation and Sorting of Pancreatic Cells
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Corresponding Organization : City of Hope
Other organizations : University of Southern California, Jawaharlal Nehru University, Royal College of Surgeons in Ireland - Bahrain, Children's Hospital of Los Angeles
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Variable analysis
independent variables
- Perfusion with the Liberase enzyme blend (Sigma Aldrich) for isolating islets from mouse pancreas
- Harvesting and digestion of pancreatic buds from Pdx1-Cre:Rosa26R-YFP embryos, heterozygous Ngn3-EGFP embryos, or the MIP-GFP transgenic pups into single cells using TrypLE Express (ThermoFisher)
- Percentage purity (85–95%) of sorted cells for green fluorescent protein imaging using a BD FACS Aria II
- Wild-type cells as the negative control for FACS gating
- Positive control: Not explicitly mentioned
- Negative control: Wild-type cells used for FACS gating
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