Talbot Neil C, Shannon Amy E, Garrett Wesley M
Agricultural Research Service, NEA, Beltsville Agricultural Research Center, Animal Biosciences and Biotechnology Laboratory, U.S. Department of Agriculture, Bldg. 200, Rm. 13, BARC-East, Beltsville, MD, 20705, USA.
In Vitro Cell Dev Biol Anim. 2019 Apr;55(4):285-301. doi: 10.1007/s11626-019-00336-5. Epub 2019 Mar 13.
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.
本文描述了一种细胞系PICM - 31D的分离过程，该细胞系具有类似胰腺导管细胞的表型特征。PICM - 31D细胞系源自先前描述的猪胚胎干细胞衍生的外分泌胰腺细胞系PICM - 31。PICM - 31D细胞系在形态上与亲代细胞不同，它以单层形式生长，而不是自组装成多细胞腺泡样结构。PICM - 31D细胞以1:3至1:10的传代比例传代培养了一年多，其表型和生长速率均无变化。电子显微镜显示这些细胞是立方体细胞组成的极化上皮细胞，在其顶端/侧面通过类似紧密连接的黏附相连。细胞在顶端细胞膜下含有大量黏液样分泌小泡。对PICM - 31D细胞蛋白质进行蛋白质组学分析检测到了MUC1和MUC4，这与黏液小泡的形态一致。基因表达分析表明，这些细胞表达胰腺导管细胞相关的转录因子，如GATA4、GATA6、HES1、HNF1A、HNF1B、ONECUT1（HNF6）、PDX1和SOX9，但几乎不表达或不表达胰腺祖细胞标志物，如PTF1A、NKX6 - 1、SOX2或NGN3。PICM - 31D细胞表达胰腺导管细胞相关基因，包括CA2、CFTR、MUC1、MUC5B、MUC13、SHH、TFF1、KRT8和KRT19，但不表达外分泌胰腺标志物基因CPA1和PLA2G1B。然而，外分泌标志物AMY2A仍在这些细胞中表达。令人惊讶的是，尿路上皮蛋白在PICM - 31D细胞蛋白质组中很突出，尤其是UPK1A。膜联蛋白A1和A2蛋白在细胞中也相对丰富。在细胞中检测到了尿路上皮蛋白和膜联蛋白基因的表达，尽管在胎猪胰腺导管组织中仅检测到UPK1B、UPK3B、ANXA2和ANXA4。总之，PICM - 31D细胞系模拟了胎猪胰腺分泌黏液的导管细胞。
