Cell Biology Core

细胞生物学核心

基本信息

批准号：
7466206
负责人：
NASSRIN DASHTI
金额：
$ 5.88万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7466206
关键词：
Address Adhesions Air Alabama American Type Culture Collection Anabolism Animals Antibiotic Resistance Antibiotics Apical Apolipoproteins Apolipoproteins A Apolipoproteins B Appendix Ascites Atherosclerosis Benzamidines Biochemical Biogenesis Biological Assay Bos taurus Breeding C-Peptide Caco-2 Cells Caliber Canis familiaris Carbon Dioxide Cardiovascular Diseases Caring Catabolism Cattle Cell Line Cells Cellular biology Characteristics Chloramphenicol Cholesterol Chylomicrons Class Clinical Colon Adenocarcinoma Complementary DNA Complex Condition Conditioned Culture Media Cultured Cells Cysteine Cytolysis DNA Diet Eagles Edetic Acid Endothelial Cells Enterocytes Equus caballus Experimental Models Fasting Fat-Free Diets Fatty acid glycerol esters Gelatin Gene Silencing Genes Glutamine Glycerol Goals Growth Factor HEPES Heating Heparin Hepatic Hepatoblastoma Hepatocyte High Density Lipoprotein Cholesterol High Density Lipoproteins Housing Human Human Cell Line IACUC Immunoblot Analysis Immunoprecipitation In Vitro Incubated Injection of therapeutic agent Intestinal Secretions Intestines Kidney Kinetics L-Glucose Label Laboratories Letters Libraries Lipids Lipoproteins Liver Liver parenchyma Low-Density Lipoproteins Lymph Maintenance Measurement Mediating Membrane Mesentery Mesocricetus auratus Metabolic Metabolism Methionine Methods MicroRNAs Modeling Mus Mutate Non-Essential Amino Acid Nutritional Oleic Acid Oleic Acids Pathway interactions Penicillin G Peptides Peritoneal Macrophages Phenylmethylsulfonyl Fluoride Phosphate Buffer Phospholipid Transfer Proteins Phospholipids Physiological Plasma Plastics Play Primary carcinoma of the liver cells Procedures Process Production Property Protein Analysis Protein Overexpression Proteins Protocols documentation Publishing Purpose Pyruvate Pyruvates RNA Interference Rattus Regulation Relative (related person)Research Design Role Saline Sampling Saphenous Vein Serum Services Side Site Sodium Sodium Bicarbonate Sodium Chloride Staging Streptomycin Sulfate (2[{..}]3) Salt Structure Structure-Activity Relationship Study models Suspension substance Suspensions Syringes Techniques Terminator Codon Testing Thioglycolates Time Transfection Translations Universities Very low density lipoprotein Week Western Blotting animal resource apolipoprotein B-48 base benzamidine calcium phosphate cholesterol absorption day density fetal flasks hexanoic acid hormone regulation in vivo interest lipid transfer protein macrophage microsomal triglyceride transfer protein mimetics monocyte monolayer multicatalytic endopeptidase complex mutant particle peptide A peptide L phospholipid exchange proteins planetary Atmosphere polycarbonate prevent programs research study size stable cell line stoichiometry tissue culture vector

项目摘要

A. STATEMENT OF OBJECTIVES The Cell Biology Core is a new facility and will serve all four projects. This core will provide several lipoprotein-producing and non-lipoprotein producing cell lines required for studies described in each project. The lipoprotein producing cells will be used: 1) To study the mechanisms of assembly and secretion of endogenous apoA-l- and apoB-containing nascent lipoprotein particles; 2) To determine the effects of several apoA-l mimetic peptides on the concentration, composition, and properties of apoA-l- and apoB-containing nascent lipoprotein particles; 3) For transfection studies to assess the structure-function relationship of apoB, and potentially apoA-l, and their respective particle assembly and secretion; 4) To silence the gene of interest using miRNA technique; 5) To establish clonal stable transformants of cells that express C-terminally truncated forms of human apoB or cells that lack specific gene, i.e, lipid transfer proteins. The non-lipoprotein producing cell lines will be used: 1) For the expression of ABCA1, ABCG1, and ABCG4; 2) To established clonal statble cell lines overexpressing ABCA1, ABCG1, and ABCG4; 3) To evaluate the effects of apoA-l mimetic peptides on LPS-induced adhesion of THP-1 monocytes to HUVECs; 4) For cholesterol efflux studies.

A. 目标声明细胞生物学核心是一个新设施，将为所有四个项目提供服务。该核心将提供研究所需的几种产生脂蛋白和不产生脂蛋白的细胞系每个项目中都有描述。脂蛋白产生细胞将用于： 1) 研究组装和组装的机制分泌内源性含apoA-l-和apoB的新生脂蛋白颗粒； 2) 至确定几种 apoA-l 模拟肽对浓度、组成和浓度的影响含有 apoA-l 和 apoB 的新生脂蛋白颗粒的特性； 3) 用于转染评估 apoB 和潜在的 apoA-l 的结构-功能关系及其它们的研究各自的颗粒组装和分泌； 4) 使用 miRNA 沉默感兴趣的基因技术; 5) 建立C末端表达细胞的克隆稳定转化体人类 apoB 的截短形式或缺乏特定基因（即脂质转移蛋白）的细胞。将使用非脂蛋白产生细胞系：1)用于ABCA1的表达， ABCG1和ABCG4； 2) 建立过表达ABCA1的克隆稳定细胞系， ABCG1和ABCG4； 3) 评估apoA-l模拟肽对LPS诱导的影响 THP-1单核细胞与HUVEC的粘附； 4) 用于胆固醇流出研究。