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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.目标说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。这个核心将提供需要研究的几个产生脂蛋白和非脂蛋白的细胞系在每个项目中都有描述。脂蛋白产生细胞将被用于：1)研究组装和分泌内源性载脂蛋白A-L和载脂蛋白B-新生脂蛋白颗粒；2) 测定几种载脂蛋白A-L模拟肽对血清中载脂蛋白A的浓度、组成和活性的影响含载脂蛋白A-L和载脂蛋白B的新生脂蛋白颗粒的性质；3)用于转染法评估载脂蛋白B和潜在的载脂蛋白A-L的结构-功能关系及其研究各自的颗粒组装和分泌；4)利用miRNA沉默目的基因技术；5)建立C末端表达细胞的克隆稳定转化子截短形式的人载脂蛋白B或缺乏特定基因的细胞，即脂转移蛋白。非脂蛋白产生细胞系将用于：1)ABCA1的表达， Abcg1和ABCG4；2)建立了高表达ABCA1的克隆稳定细胞系， 3)观察载脂蛋白A-L模拟肽对内毒素诱导的内毒素诱导的影响 THP-1单核细胞与人脐静脉内皮细胞的黏附；4)胆固醇外流研究。