Talbot Neil C, Shannon Amy E, Phillips Caitlin E, Garrett Wesley M
From the Animal Biosciences and Biotechnology Laboratory, Agricultural Research Service, US Department of Agriculture, Beltsville, MD.
Pancreas. 2017 Jul;46(6):789-800. doi: 10.1097/MPA.0000000000000836.
The aim of this study was to identify an epithelial cell line isolated from the spontaneous differentiation of totipotent pig epiblast cells.
PICM-31 and its colony-cloned derivative cell line, PICM-31A, were established from the culture and differentiation of an epiblast mass isolated from an 8-day-old pig blastocyst. The cell lines were analyzed by transmission electron microscopy, marker gene expression, and mass spectroscopy-based proteomics.
The PICM-31 cell lines were continuously cultured and could be successively colony cloned. They spontaneously self-organized into acinarlike structures. Transmission electron microscopy indicated that the cell lines' cells were epithelial and filled with secretory granules. Candidate gene expression analysis of the cells showed an exocrine pancreatic profile that included digestive enzyme expression, for example, carboxypeptidase A1, and expression of the fetal marker, α-fetoprotein. Pancreatic progenitor marker expression included pancreatic and duodenal homeobox 1, NK6 homeobox 1, and pancreas-specific transcription factor 1a, but not neurogenin 3. Proteomic analysis of cellular proteins confirmed the cells' production of digestive enzymes and showed that the cells expressed cytokeratins 8 and 18.
The PICM-31 cell lines provide in vitro models of fetal pig pancreatic exocrine cells. They are the first demonstration of continuous cultures, that is, cell lines, of nontransformed pig pancreas cells.
本研究旨在鉴定一种从全能猪上胚层细胞自发分化分离得到的上皮细胞系。
PICM - 31及其集落克隆衍生细胞系PICM - 31A，是从8日龄猪囊胚分离的上胚层团块的培养和分化过程中建立的。通过透射电子显微镜、标记基因表达和基于质谱的蛋白质组学对这些细胞系进行分析。
PICM - 31细胞系能够连续培养，并可连续进行集落克隆。它们能自发地自我组织形成腺泡样结构。透射电子显微镜显示，该细胞系的细胞为上皮细胞，充满分泌颗粒。对这些细胞的候选基因表达分析显示出外分泌胰腺特征，包括消化酶表达，如羧肽酶A1，以及胎儿标志物甲胎蛋白的表达。胰腺祖细胞标志物表达包括胰腺和十二指肠同源盒1、NK6同源盒1和胰腺特异性转录因子1a，但不包括神经生成素3。对细胞蛋白质的蛋白质组学分析证实了这些细胞产生消化酶，并表明这些细胞表达细胞角蛋白8和18。
PICM - 31细胞系提供了胎猪胰腺外分泌细胞的体外模型。它们是未转化猪胰腺细胞连续培养即细胞系的首次证明。
