Supplementary MaterialsSupplementary Physique 1: Human pancreatic development 1. years old (A) and 10 years aged (B), stained by immunohistochemistry for Insulin (pink), Glucagon (blue), and Ki67 (brown) with a hematoxylin counterstain. Insets, high power images of the indicated area marked by black squares in the low power images. Scale bars, 200 m in low power images and 100 m in insets. Image_3.jpg (2.5M) GUID:?EB7C4621-2689-4DA8-B956-EC2F93A2F7EE Supplementary Physique 4: A rare example of replicating chromograninA positive hormone-negative (CPHN) cells in a fetal and an infant donor. Pancreatic sections from a fetal (A) and an infant (B) donor immunostained for Endocrine cocktail (insulin, glucagon, somatostatin, pancreatic polypeptide, and ghrelin) (white), chromograninA (green), Ki67 (reddish), and DAPI (blue). Yellowish arrows displaying Ki67 positive CPHN cells in a single one and fetal baby donor, emphasizing that replication is really a uncommon event in these cells. Range pubs: 100 m for low power and 25 m for high magnification pictures. Picture_4.jpg (1.2M) GUID:?D7D117F9-9D72-421A-A0A0-E555957F3F1E Supplementary Body 5: Replication and expression of pan-endocrine hormones in cells within the ducts and PDGs of fetal and infant pancreas. Representative pancreatic areas from fetal Rigosertib sodium and baby donors stained for Ki67/Hematoxylin (A,B, respectively) and Insulin/PP/hematoxylin (C,D, respectively). Insets, higher magnification of chosen areas (indicated by dark squares) in the reduced power pictures. Dark brown arrows (within a,B and their insets) suggest Ki67 staining (replication of cells) in ducts and PDGs. Dark brown arrows (insets of C,D) suggest appearance of pancreatic polypeptide (PP) and crimson arrows indicate appearance of insulin in PDGs. Range pubs, 100 m (for the,B), 200 m (for C,D), 25 m (for all your insets). Picture_5.jpg (2.4M) GUID:?3F759301-B294-4E82-AFC1-71D1DA6BACAB Supplementary Body 6: Chromogranin A confident hormone-negative (CPHN) cells situated in the pancreatic ducts usually do not replicate during fetal and baby lifestyle. Pancreatic ducts proven in tissue areas from fetal (A) and baby (B) donors immunostained for Endocrine cocktail (insulin, glucagon, somatostatin, pancreatic polypeptide and ghrelin) (white), chromograninA (green), Ki67 (crimson), and DAPI (blue). Yellow arrows show CPHN cells. Level bars: 100 m for low power and 25 m for high magnification images. Image_6.jpg (1.0M) GUID:?DCBE1900-772B-444D-B546-7581005D6D25 Supplementary Figure 7: Replication of endocrine cells. Quantification of endocrine cell replication demonstrated as percentage of Ki67 positive endocrine cells, immunostained with endocrine cocktail antibodies. Endocrine cell replication diminishes in the pancreas with age ( 0.05). Image_7.jpg (84K) GUID:?D5BBD777-1E2D-4E9A-913E-CB83A685D05F Supplementary Number 8: Examples of replicating islet endocrine cells inside a fetal and an infant donor. Pancreatic sections from a fetal (A) and an infant (B) donor immunostained for Endocrine cocktail (insulin, glucagon, somatostatin, pancreatic polypeptide and ghrelin) (white), chromograninA (green), Ki67 (reddish), and DAPI (blue). Yellow arrows showing Ki67 positive endocrine cells in high power images indicated by reddish squares in low power images. The percentage of replication of islet endocrine cells decreased from fetal to postnatal existence Rigosertib sodium (C). Scale bars: 50 m for low power and 10 m for high magnification images. Image_8.jpg (1.2M) GUID:?579F131B-5E6C-4892-B01E-301F54FBA0C0 Supplementary Figure 9: Percent changes of CPHN cells (positive for either NKx6.1 or NKx2.2) in different compartments of fetal and infant/child pancreas with age: The percentage of either NKX6.1+ or NKX2.2+ CPHN cells (of total CPHN cells in fetal and infant/child instances) found in overall compartments (A,E), within islets (B,F), in cluster cells (C,G) or in solitary cells Rigosertib sodium (D,H). Image_9.jpg (571K) GUID:?42373E2A-BBB4-4E8F-A0EC-E8237B33E29B Supplementary Table 1: Clinical characteristic of fetal and infant cases used for quantification of CPHN cells. PT, pancreas tail. Table_1.DOCX (77K) GUID:?FEC59B3A-CC8B-4302-8DF7-BAE50A6F8FD5 Supplementary Table 2: Clinical characteristics of nPOD fetal and infant donors for Ki67, Nkx2.2 and Nkx6.1 analysis. PH, pancreas head; PB, pancreas body; PT, pancreas tail. Table_2.DOCX (99K) GUID:?A06E3EF1-801D-4B14-9E44-2A31A79F531A Supplementary Table 3: Clinical characteristics of nPOD fetal and infant instances for Ki67 and hormone expression analysis in pancreatic ducts. PH, pancreas head; PB, pancreas body; PT, pancreas tail. Table_3.DOCX (101K) GUID:?D769E7BA-F79B-4E7E-A8A8-675E4B16FDAE Supplementary Table 4: NKX6.1 + and NKX2. 2 + CPHN cells recognized in differentcompartments Rabbit Polyclonal to CARD11 of the pancreas in fetal and infant donors. Table_4.DOCX (85K) GUID:?64F41419-1E3F-40CD-A81F-3CBA14944E2F Abstract Context: Previously, we identified chromograninA positive hormone-negative (CPHN) cells in high frequency in human being fetal and neonatal pancreas, likely representing nascent endocrine precursor cells. Here, we characterize the putative endocrine fate and replicative status of these newly created cells. Objective: To establish the replicative rate of recurrence and transcriptional identity of CPHN cells, extending our observation on CPHN cell rate of recurrence to a larger cohort of fetal and infant pancreas. Design, Setting, and Participants: 8 fetal, 19 infant autopsy pancreata were evaluated for CPHN cell rate of recurrence; 12 fetal, 24 infant/child.