Nielsen Michael Friberg Bruun, Mortensen Michael Bau, Detlefsen Sönke
Department of Pathology, Odense University Hospital, J.B. Winsløws Vej 15, 5000, Odense C, Denmark.
Department of Clinical Research, University of Southern Denmark, J.B. Winsløws Vej 19, 5000, Odense C, Denmark.
Histochem Cell Biol. 2017 Oct;148(4):359-380. doi: 10.1007/s00418-017-1581-5. Epub 2017 May 25.
Pancreatic stellate cells (PSCs) play a central role as source of fibrogenic cells in pancreatic cancer and chronic pancreatitis. In contrast to quiescent hepatic stellate cells (qHSCs), a specific marker for quiescent PSCs (qPSCs) that can be used in formalin-fixed and paraffin embedded (FFPE) normal human pancreatic tissue has not been identified. The aim of this study was to identify a marker enabling the identification of qPSCs in normal human FFPE pancreatic tissue. Immunohistochemical (IHC), double-IHC, immunofluorescence (IF) and double-IF analyses were carried out using a tissue microarray consisting of cores with normal human pancreatic tissue. Cores with normal human liver served as control. Antibodies directed against adipophilin, α-SMA, CD146, CRBP-1, cytoglobin, desmin, GFAP, nestin, S100A4 and vinculin were examined, with special emphasis on their expression in periacinar cells in the normal human pancreas and perisinusoidal cells in the normal human liver. The immunolabelling capacity was evaluated according to a semiquantitative scoring system. Double-IF of the markers of interest together with markers for other periacinar cells was performed. Moreover, the utility of histochemical stains for the identification of human qPSCs was examined, and their ultrastructure was revisited by electron microscopy. Adipophilin, CRBP-1, cytoglobin and vinculin were expressed in qHSCs in the liver, whereas cytoglobin and adipophilin were expressed in qPSCs in the pancreas. Adipophilin immunohistochemistry was highly dependent on the preanalytical time interval (PATI) from removal of the tissue to formalin fixation. Cytoglobin, S100A4 and vinculin were expressed in periacinar fibroblasts (FBs). The other examined markers were negative in human qPSCs. Our data indicate that cytoglobin and adipophilin are markers of qPSCs in the normal human pancreas. However, the use of adipophilin as a qPSC marker may be limited due to its high dependence on optimal PATI. Cytoglobin, on the other hand, is a sensitive marker for qPSCs but is expressed in FBs as well.
胰腺星状细胞（PSC）在胰腺癌和慢性胰腺炎中作为纤维化细胞的来源发挥着核心作用。与静止的肝星状细胞（qHSC）不同，尚未鉴定出可用于福尔马林固定石蜡包埋（FFPE）正常人胰腺组织的静止PSC（qPSC）的特异性标志物。本研究的目的是鉴定一种能够在正常人FFPE胰腺组织中鉴定qPSC的标志物。使用由正常人胰腺组织芯组成的组织微阵列进行免疫组织化学（IHC）、双重IHC、免疫荧光（IF）和双重IF分析。正常人肝脏芯用作对照。检测了针对脂联素、α-SMA、CD146、CRBP-1、细胞珠蛋白、结蛋白、GFAP、巢蛋白、S100A4和纽蛋白的抗体，特别关注它们在正常人胰腺腺泡周围细胞和正常人肝脏窦周细胞中的表达。根据半定量评分系统评估免疫标记能力。将感兴趣的标志物与其他腺泡周围细胞的标志物一起进行双重IF。此外，还检测了组织化学染色在鉴定人qPSC中的效用，并通过电子显微镜重新观察了它们的超微结构。脂联素、CRBP-1、细胞珠蛋白和纽蛋白在肝脏的qHSC中表达，而细胞珠蛋白和脂联素在胰腺的qPSC中表达。脂联素免疫组织化学高度依赖于从组织切除到福尔马林固定的分析前时间间隔（PATI）。细胞珠蛋白、S100A4和纽蛋白在腺泡周围成纤维细胞（FB）中表达。其他检测的标志物在人qPSC中呈阴性。我们的数据表明，细胞珠蛋白和脂联素是正常人胰腺中qPSC的标志物。然而，由于脂联素对最佳PATI的高度依赖性，其作为qPSC标志物的应用可能受到限制。另一方面，细胞珠蛋白是qPSC的敏感标志物，但也在FB中表达。
