Fig. 2. Human pancreatic slices remain viable after extended culture.

a Representative immunostaining showing tissue viability at the time of tissue slicing, showing live (green) and dead (red) regions. Scale: 500 µm for all panels. b Immunostaining of control fixed slice showing only dead cells. Scale: 500 µm. c Immunostaining showing slice viability assessments after 10 days of culture in either transwells or PFC dishes. Necrotic zones in transwell-cultured slices are highlighted in magnified panel. Overt necrosis, in contrast, is not apparent in PFC-cultured HPSs. Scale: 500 µm. Magnified insets: 200 µm. For a–c: n = 5 biologically independent samples from individual donors. d Quantitative measurement of viable cell percentages in PFC (red)- and transwell (blue)-cultured slices over a period of 10 days. Shaded area shows mean ± SD. e Bar plots showing cellular viability of either culture setting at day 5 and 10 post-slicing. Two-tailed t-test: **p-val_10days = 0.002. f Slice cross-sectional area of slices kept in either culture condition for 10 days in culture. Shaded area shows mean ± SD. g Individual bar plots showing slice cross-sectional area at days 7, 14, and 21 post slicing. Two-tailed t-test: **p-val_14days = 0.0036, ****p-val_21days = 0.000087. For d–g: n = 4 biologically independent samples from individual donors. h Viable cell percentages of PFC- and transwell-cultured T1D donor slices over a period of 10 days (n = 1 sample from an individual donor). i Slice cross-sectional area in either culture condition over 10 days of culture (T1D donor) (n = 1 sample from an individual donor). j Immunofluorescence images showing expression of representative endocrine markers (INS insulin, white; GCG glucagon, red; and SST somatostatin, green) within islets of slices at day 0 and cultured for 10 days in either transwells or PFC dishes. Scale for all panels: 500 µm; 100 µm for insets. k Dot plot showing the distribution of endocrine cells in islets at day 0 and after 10 days of culture (PFC and transwells). l Dot plot showing the cross-sectional area of islets at day 0 and after 10 days of culture (PFC and transwells). Two-tailed t-test: *p-val_{D10 (PFC)-D10 (TW)} = 0.0107. For j–l: data from n = 10 biologically independent samples from individual donors. m Immunostaining showing insulin (INS, white) and representative exocrine markers (cytokeratin-19, CK-19, red; and amylase, AMY, green) within slices at day 0 and after 10 days of culture (PFC and transwells). Blue, DAPI counter-staining. n = 10 biologically independent samples from individual donors. Scale: 500 µm; 100 µm for insets. For d–i, k, and l, data are presented as mean ± SD. Each n further represents the mean of three technical replicates. * p < 0.05; **p < 0.01; ***p < 0.005. Source are provided in the Source Data file.