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新的手段。然而，使用该数据库鉴定新的诊断/预后标志物和定义疾病的分子机制的限制因素是从临床组织活检中分离特定细胞群及其mRNA的挑战。然而，它们需要高度的手工技能并且耗时。NIH开发了激光捕获显微切割（LCM）系统，以应对这一挑战，并更快速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- 1，000个细胞中产生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LCM设施将并入埃默里大学血管医学中心。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，干细胞移植后的移植物抗宿主病，以及关于慢性肝病期间体内第八因子来源的研究。将利用LCM设施的具体程序包括：1）比较对照患者和慢性丙型肝炎患者肝活检样本中Kupffer细胞、内皮星状细胞和T细胞内的基因表达（急性vs.慢性;非进展性vs.重度进展性炎症/纤维化的同胞;治疗应答者vs.治疗无应答者）; 2）研究暴露于不同血流动力学环境和治疗的人和实验动物中个体内皮细胞中基因表达的调节; 3）研究血管成形术后再狭窄期间血管外膜肌成纤维细胞的体内基因表达; 4）研究血管成形术后再狭窄期间血管外膜肌成纤维细胞的体内表达; 4）研究慢性肝病期间肝内皮和实质细胞中因子VIII的基因表达; 5）研究增殖与消退血管瘤的内皮细胞的体内基因表达;和6）研究皮肤炎性疾病期间人真皮内皮细胞的体内基因表达。

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.