Pasternak Artur, Gajda Mariusz, Gil Krzysztof, Matyja Andrzej, Tomaszewski Krzysztof A, Walocha Jerzy A, Kulig Jan, Thor Piotr
First Department of General, Oncological and Gastrointestinal Surgery, Jagiellonian University Medical College, Krakow, Poland.
Folia Histochem Cytobiol. 2012;50(4):581-5. doi: 10.5603/19673.
The aim of this study was to assess the presence of interstitial Cajal-like cells (ICLCs) in the human gallbladder and to determine their distinctive characteristics on the basis of double immunohistochemical staining. Gallbladder specimens were obtained from 30 patients subjected to elective laparoscopic cholecystectomy for symptomatic gallstone disease. Tissue samples were fixed in 4% phosphate-buffered paraformaldehyde, processed, embedded in paraffin, and, after sectioning, routinely stained with HE. Tissue antigens were retrieved using the heat-induced epitope retrieval (HIER) method. For simultaneous visualisation of two antigens, an indirect double immunofluorescence procedure was applied. ICLCs were defined as CD117-immunopositive and tryptase-immunonegative objects. They were predominantly fusiform in shape with sparse branches that were visible in some sections. ICLCs were observed throughout the organ including the gallbladder's fundus, body (corpus) and neck, being most numerous in the corpus. The ICLCs were detected almost exclusively within the muscularis propria and they were arranged parallel to smooth muscle cells. The following subpopulations of ICLCs were observed: ICLC-IM (intramuscular ICLCs) localised between smooth muscle fibres forming one muscle bundle; and ICLC-IB (interbundle ICLCs) localised within the connective tissue separating smooth muscle bundles. Thus, the presence of ICLCs in the human gallbladder was clearly identified, demonstrated by double immunohistochemistry which was found to be a reliable method for differentiating ICLCs from mast cells.
本研究旨在评估人胆囊中间质 Cajal 样细胞(ICLCs)的存在情况,并通过双重免疫组化染色确定其独特特征。胆囊标本取自 30 例因有症状胆结石疾病接受择期腹腔镜胆囊切除术的患者。组织样本用 4%磷酸盐缓冲多聚甲醛固定,经过处理后包埋于石蜡中,切片后常规进行 HE 染色。采用热诱导抗原修复(HIER)方法对组织抗原进行修复。为同时可视化两种抗原,应用间接双重免疫荧光法。ICLCs 被定义为 CD117 免疫阳性且类胰蛋白酶免疫阴性的细胞。它们主要呈梭形,在某些切片中可见稀疏分支。在整个胆囊器官包括胆囊底部、体部(胆囊体)和颈部均观察到 ICLCs,其中以胆囊体部数量最多。ICLCs 几乎仅在固有肌层内被检测到,且它们与平滑肌细胞平行排列。观察到以下 ICLCs 亚群:ICLC-IM(肌内 ICLCs)位于形成一个肌束的平滑肌纤维之间;以及 ICLC-IB(束间 ICLCs)位于分隔平滑肌束的心结缔组织内。因此,通过双重免疫组化明确鉴定出了人胆囊中 ICLCs 的存在,该方法被发现是区分 ICLCs 与肥大细胞的可靠方法。