Emory Medicine Laser Capture Microdissection Facility

人类基因组数据库中的信息有望为研究人类疾病提供一种新的手段。然而，在使用这个数据库来识别新的诊断/预后标记和确定疾病的分子机制方面，一个限制因素是从临床组织活检中分离特定细胞群体及其mRNA的挑战，研究人员已经使用微管解剖方法从小样本组织中分离出特定细胞类型。然而，它们需要很高的手工技能，而且很耗时。美国国立卫生研究院开发了激光捕获显微切割(LCM)系统，以应对这一挑战，并更快地从临床活检样本中分离特定细胞。通过将LCM与非嗜热RNA扩增方法相结合，现在可以从100-1000个细胞中生产ArNA，并探测高密度阵列，用于临床活检中特定细胞的体内功能基因组研究。埃默里医学公司的LCM设施将并入埃默里大学的血管医疗中心。它将用于研究各种慢性炎症性疾病中特定细胞类型的基因表达，如动脉粥样硬化、丙型肝炎、干细胞移植后移植物对宿主疾病，以及关于演员在慢性肝病期间的体内来源的研究。将利用LCM设备的具体项目包括：1)比较对照组患者和慢性丙型肝炎患者(急性和慢性；非进展性炎症/纤维化同胞和严重进展性炎症/纤维化患者；治疗应答者和无应答者)肝活检样本中Kupffer、内皮星状细胞和T细胞的基因表达；2)研究不同血流动力学环境和治疗方法对人和实验动物单个内皮细胞基因表达的调节；3)研究血管成形术后再狭窄过程中血管外膜成纤维细胞的基因表达；4)血管成形术后再狭窄过程中血管外膜肌成纤维细胞的体内表达；4)研究慢性肝病时肝内皮细胞和实质细胞中因子VIII的基因表达；5)研究血管瘤增生期和消退期内皮细胞的基因表达；6)皮肤炎性疾病中人真皮内皮细胞的基因表达。

The information in the human genome database promises to provide a new means to investigate human diseases. However, a limiting factor in using this database in identifying new diagnostic/prognostic markers and defining the molecular mechanisms of disease is the challenge of isolating specific cell populations and their mRNA from clinical tissue biopsies Investigators have isolated specific cell types from small samples of tissue using micropipette dissection methods. However, they require a high degree of manual skill and are time-consuming. A Laser Capture Microdissection (LCM) system was developed at NIH to meet this challenge and more rapidly isolate specific cells from clinical biopsy samples. By combining LCM with non-thermophilic RNA amplification methods it is now possible to produce aRNA from 100-1,000 cells and probe high-density arrays for in vivo functional genomic studies of specific cells in clinical biopsies. The Emory Medicine LCM facility will be incorporated into the Vascular Medical Center at Emory University. It will be used to investigate gene expression of specific cell types in a variety of chronic inflammatory diseases such as atherosclerosis, hepatitis C, graft vs. host disease following stem cell transplantation, and studies regarding the in vivo source of actor VIII during chronic liver disease. Specific programs that will utilize the LCM facility include: 1) Comparing gene expression within Kupffer, endothelial stellate, and T cells in liver biopsy samples of control patients and those with chronic hepatitis C (acute vs. chronic; siblings with non-progressive vs. severe progressive inflammation/fibrosis; responders vs. non-responders to treatment); 2) Studying the modulation of gene expression in individual endothelial cells in humans and experimental animals exposed to different hemodynamic milieus and treatments; 3) Studying the in vivo gene expression of vascular adventitial myofibroblasts during post- angioplasty restenosis; 4) Studying the in vivo expression of vascular adventitial myofibroblasts during post-angioplasty restenosis; 4) Studying gene expression of Factor VIII in liver endothelial and parenchyma cells during chronic liver diseases; 5) studying the in vivo gene expression of endothelial cells of proliferating vs. regressing hemangiomas; and 6) studying the in vivo gene expression of human dermal endothelial cells during cutaneous inflammatory diseases.