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Pancreatic duct-like cell line derived from pig embryonic stem cells: expression of uroplakin genes in pig pancreatic tissue

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Abstract

The isolation of a cell line, PICM-31D, with phenotypic characteristics like pancreatic duct cells is described. The PICM-31D cell line was derived from the previously described pig embryonic stem cell-derived exocrine pancreatic cell line, PICM-31. The PICM-31D cell line was morphologically distinct from the parental cells in growing as a monolayer rather than self-assembling into multicellular acinar-like structures. The PICM-31D cells were propagated for over a year at split ratios of 1:3 to 1:10 at each passage without change in phenotype or growth rate. Electron microscopy showed the cells to be a polarized epithelium of cuboidal cells joined by tight junction-like adhesions at their apical/lateral aspect. The cells contained numerous mucus-like secretory vesicles under their apical cell membrane. Proteomic analysis of the PICM-31D’s cellular proteins detected MUC1 and MUC4, consistent with mucus vesicle morphology. Gene expression analysis showed the cells expressed pancreatic ductal cell-related transcription factors such as GATA4, GATA6, HES1, HNF1A, HNF1B, ONECUT1 (HNF6), PDX1, and SOX9, but little or no pancreas progenitor cell markers such as PTF1A, NKX6-1, SOX2, or NGN3. Pancreas ductal cell-associated genes including CA2, CFTR, MUC1, MUC5B, MUC13, SHH, TFF1, KRT8, and KRT19 were expressed by the PICM-31D cells, but the exocrine pancreas marker genes, CPA1 and PLA2G1B, were not expressed by the cells. However, the exocrine marker, AMY2A, was still expressed by the cells. Surprisingly, uroplakin proteins were prominent in the PICM-31D cell proteome, particularly UPK1A. Annexin A1 and A2 proteins were also relatively abundant in the cells. The expression of the uroplakin and annexin genes was detected in the cells, although only UPK1B, UPK3B, ANXA2, and ANXA4 were detected in fetal pig pancreatic duct tissue. In conclusion, the PICM-31D cell line models the mucus-secreting ductal cells of the fetal pig pancreas.

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Acknowledgments

The authors thank Ms. Caitlin Phillips for her assistance in primer design and for preliminary RT-PCR analysis of the PICM-31D cell line.

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Correspondence to Neil C. Talbot.

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Care and treatment of pigs in this study were approved by the Institutional Animal Care and Use Committee of the U.S. Department of Agriculture, Beltsville Agricultural Research Center, Beltsville, MD.

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The authors declare that they have no conflict of interest.

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Editor: Tetsuji Okamoto

Mention of trade names or commercial products in this publication is solely for the purposes of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.

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Supplementary Figure 1

Semi-quantitative RT-PCR assay of the expression of pancreas duct cell genes and uroplakin genes in fetal pig pancreas tissue (FP), weanling pig (21-d old) pancreas tissue (WP), fetal pig intestinal tissue (FI), and weanling pig intestinal tissue (WI). Weanling pig intestine mRNA was used for the -RT control. (PNG 210 kb)

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Supplementary Table 1

(DOCX 18 kb)

ESM 1

Supplementary Data Sheet 1 (DOCX 862 kb)

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Talbot, N.C., Shannon, A.E. & Garrett, W.M. Pancreatic duct-like cell line derived from pig embryonic stem cells: expression of uroplakin genes in pig pancreatic tissue. In Vitro Cell.Dev.Biol.-Animal 55, 285–301 (2019). https://doi.org/10.1007/s11626-019-00336-5

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  • DOI: https://doi.org/10.1007/s11626-019-00336-5

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