结肠粘膜活检标本中固有层单个核细胞的分离及表型分析

Isolation and Phenotypic Analysis of Lamina Propria Mononuclear Cells from Colonoscopic Biopsy Specimens

为改进从粘膜活检标本中分离固有层单个核细胞的方法 ,并行初步表型分析 ,从 8例溃疡性结肠炎、5例正常人和 2 2例大肠癌患者各取 1 0块结肠粘膜进行活检 ,比较消化分离各因素对细胞产量的影响 ,并以双色荧光流式细胞术分析淋巴细胞各亚群的变化。结果显示 :溃疡性结肠炎组固有层单个核细胞产量达 1 0 6 个 ,为正常人组及大肠癌组的 2倍 ,各组细胞活力均 >95 % ;其中 T、B细胞百分比在三组间无统计学上的差异 ,而溃疡性结肠炎组 CD3+ CD4+ 、CD3+ CD8+ T细胞百分比与后两组比较则有显著不同 (P<0 .0 1 )。本研究结果提示 ,从活检标本中可分离出足够数量的固有层单个核细胞进行表型或功能研究 。

This study was directed to the method of isolation of lamina propria mononuclear cells (LPMC) from mucosal biopsy specimens and the phenotypic analysis of the cells. Ten biopsy specimens, taken individually from 8 patients with ulcerative colitis (UC), 5 normal controls and 22 patients with colorectal cancer, were collected to compare the factors that influence isolation and cell yield and to analyze the lymphocyte subsets by means of two color flow cytometry. LPMC yield averaged 10 6 with a viability of more than 95% in UC, twice the yields in other two groups. The percentages of total T, B cells in three groups were similar ( P >0.05). When the UC group was compared with the other two groups, a significantly higher proportion of CD 3 + CD 4 + T cell (49 98% vs 37.54% and 37.25%, P <0.01) was noted, whereas a lower proportion of CD 3 + CD 8 + T cell (23 64% vs 31 52% and 31.07%, P <0.01) was observed. These data showed that the yield and viability of LPMC isolated from biopsy specimens were good enough for a phenotypic or functional analysis, which might be helpful to mucosal immune research on gut disorders, such as UC.