Molnár B, Kármán J, Németh A, Prónai L, Zágoni T, Tulassay Z
II. Belgyógyászati Klinika, Semmelweis Egyetem, Budapest.
Orv Hetil. 2000 Apr 9;141(15):789-92.
Aim of the present work was the development of a mechanic cell separation protocol for gastrointestinal biopsy analysis. Evaluation of the technique was performed on selected group of patients who underwent routine endoscopy. Routine gastrointestinal biopsies were obtained after informed consent. 23 gastric (6 healthy, 14 gastritis, 3 adenocarcinoma) and 15 colon samples (5 healthy, 7 colitis ulcerosa, 3 adenocarcinoma) were evaluated. The mechanic disruption of the biopsies was performed by Medimachine (DAKO, Denmark), a commercially available system using a 30 microns miner and a 30 microns mesh. The cell solution was centrifuged for 5 minutes by 250 g. The cells were fixed in paraformaldehide and stained by propidium iodide. The flow cytometry analysis was performed on a BD FacStar Plus flow cytometer. The DNA data were evaluated using the Winlist software. All of the preparations were appropriate for flow cytometric analysis. The coefficient of variation of the DNA histograms (n = 7) (CV mean +/- SD. 6.45% +/- 1.21) were acceptable for analysis. In the gastric biopsy samples aneuploidy was determined only in malignant cases. In four of the seven colitis ulcerosa samples and in one of the three adenocarcinoma aneploidy was found. The histologically healthy specimen were all diploid. Mechanic cell separation and disaggregation is a useful method for preparing fresh biopsy specimen for flow cytometry.
本研究的目的是开发一种用于胃肠活检分析的机械细胞分离方案。对接受常规内镜检查的特定患者群体进行了该技术的评估。在获得知情同意后获取常规胃肠活检样本。对23份胃样本(6份健康样本、14份胃炎样本、3份腺癌样本)和15份结肠样本(5份健康样本、7份溃疡性结肠炎样本、3份腺癌样本)进行了评估。活检样本的机械破碎通过Medimachine(丹麦达科公司)进行,这是一种市售系统,使用30微米的研磨器和30微米的滤网。细胞溶液以250 g离心5分钟。细胞用多聚甲醛固定,并用碘化丙啶染色。在BD FacStar Plus流式细胞仪上进行流式细胞术分析。使用Winlist软件评估DNA数据。所有制剂均适合进行流式细胞术分析。DNA直方图的变异系数(n = 7)(CV均值±标准差,6.45%±1.21)可接受用于分析。在胃活检样本中,仅在恶性病例中检测到非整倍体。在7份溃疡性结肠炎样本中的4份以及3份腺癌样本中的1份中发现了非整倍体。组织学上健康的标本均为二倍体。机械细胞分离和解聚是一种用于为流式细胞术制备新鲜活检标本的有用方法。