激光显微切割技术分离淋巴结血管内皮细胞

Separation of endothelial cell from lymph node of lymphoma using laser capture microdissection

目的探讨淋巴瘤发生的分子机制及有关的基因改变,研究分离、分析淋巴结中血管内皮细胞基因表达差异的方法。方法将淋巴瘤淋巴结标本液氮速冻,冷冻切片,快速免疫组化标记血管内皮细胞,使用激光显微切割技术分离血管淋巴细胞,提取血管内皮细胞中的 mRNA,使用 RT-PCR 检测所得 RNA 的质量。结果成功地实现了血管内皮细胞的分离,并保持了血管内皮细胞中 RNA 的完整性。结论确立了一个适于分离淋巴瘤血管内皮细胞的激光显微切割方法。

Objective To elucidate the molecular mechanism of lymphoma,and to optimize a method to study the gene expression difference of vascular endothelial cell.Methods The samples were collected and flash frozen quickly to maitaince the integrity of mRNA.Then the samples were sectioned on a cryostat. Sections were quickly stained with anti-CD_(34)antibody,and endothelial cell was captured.Then mRNA from endothelial cell of lymphoma and reactive lymph node were extracted.After been reverse transcripted,cDNA were tested for its and quanlity.Results Using zinc-fix as fixtive,we successfully separate endothelial cell from lymph node and obtained mRNA with satisfied integrity and purity.Conclusion Laser capture microdis- section is a suitable method to isolate endothelial cell from lymphoma patient.