Smooth muscle cells (SMCs) play an important role in physiology and production in farm animals such as pigs. Here, we report the generation of a pig SMC line. Our original objective was to establish an enteroendocrine cell line from the pig ileum epithelium through lentiviral transduction of the Simian Virus (SV) 40 large T antigen. However, an initial expression analysis of marker genes in nine cell clones revealed that none of them were enteroendocrine cells or absorptive enterocytes, goblet cells, or Paneth cells, some of the major cell types existing in the ileum epithelium. A more detailed characterization of one clone named PIC7 by RNA-seq showed that these cells expressed many of the known smooth muscle-specific or -enriched genes, including smooth muscle actin alpha 2, calponin 1, calponin 3, myosin heavy chain 11, myosin light chain kinase, smoothelin, tenascin C, transgelin, tropomyosin 1, and tropomyosin 2. Both quantitative PCR and RNA-seq analyses showed that the PIC7 cells had a high expression of mRNA for smooth muscle actin gamma 2, also known as enteric smooth muscle actin. A Western blot analysis confirmed the expression of SV40 T antigen in the PIC7 cells. An immunohistochemical analysis demonstrated the expression of smooth muscle actin alpha 2 filaments in the PIC7 cells. A collagen gel contraction assay showed that the PIC7 cells were capable of both spontaneous contraction and contraction in response to serotonin stimulation. We conclude that the PIC7 cells are derived from an enteric SMC from the pig ileum. These cells may be a useful model for studying the cellular and molecular physiology of pig enteric SMCs. Because pigs are similar to humans in anatomy and physiology, the PIC7 cells may be also used as a model for human intestinal SMCs.

中文翻译：

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从猪回肠产生肠平滑肌细胞系。


平滑肌细胞 (SMC) 在猪等农场动物的生理和生产中发挥着重要作用。在这里，我们报告了猪 SMC 系的产生。我们最初的目标是通过慢病毒转导猿猴病毒 (SV) 40 大 T 抗原，从猪回肠上皮建立肠内分泌细胞系。然而，对九个细胞克隆中标记基因的初步表达分析表明，它们都不是肠内分泌细胞或吸收性肠上皮细胞、杯状细胞或潘氏细胞（回肠上皮中存在的一些主要细胞类型）。通过 RNA-seq 对名为 PIC7 的克隆进行更详细的表征表明，这些细胞表达许多已知的平滑肌特异性或富集基因，包括平滑肌肌动蛋白 α 2、钙调蛋白 1、钙调蛋白 3、肌球蛋白重链 11、肌球蛋白轻链激酶、smoothelin、tenascin C、transgelin、原肌球蛋白 1 和原肌球蛋白 2。定量 PCR 和 RNA-seq 分析均显示 PIC7 细胞高表达平滑肌肌动蛋白 γ 2（也称为肠平滑肌）的 mRNA肌动蛋白。蛋白质印迹分析证实了 PIC7 细胞中 SV40 T 抗原的表达。免疫组织化学分析证明了 PIC7 细胞中平滑肌肌动蛋白 α2 丝的表达。胶原凝胶收缩测定表明，PIC7 细胞能够自发收缩和响应血清素刺激而收缩。我们得出结论，PIC7 细胞源自猪回肠的肠 SMC。这些细胞可能是研究猪肠道 SMC 细胞和分子生理学的有用模型。由于猪在解剖学和生理学上与人类相似，因此PIC7细胞也可以用作人类肠道平滑肌细胞的模型。