目的:研究胞外基质(ECM)材料木糖葡聚糖(XG)对人肝癌细胞系HepG2细胞在细胞培养板上的贴壁性及对微囊化HepG2细胞的清蛋白合成速率、NH_4~+清除速率、细胞增殖速率和间隙连接蛋白(Cx32)、细胞黏附分子(E-cadherin)表达的影响。方法:将HepG2细胞接种于不同浓度XG(0,0.5,1,4,6mg/ml)包被的细胞培养板上,孵育4h后收集贴壁细胞,BCA法测贴壁细胞总蛋白量进而测定细胞贴壁率;利用自制装置制备包被HepG细胞的海藻酸-多聚赖氨酸-海藻酸(APA)微囊并在培养微囊第6,10,14天以Human Serum Albumin Elisa Kit、Ammonia Assay Kit分别测定培养液中人血清白蛋白(HSA)和处理溶液中NH_4~+的浓度,检测微囊化HepG2细胞的清蛋白合成速率和NH_4~+清除速率;微囊培养1,2,3d后裂解微囊收集HepG2细胞并以RT-PCR方法检测细胞内Cx32和E-cadherin的表达情况。结果:HepG2细胞在XG包被的细胞培养板上的贴壁率随XG浓度增大呈递增趋势,同时微囊内细胞数随XG浓度增大、培养时间延长而增加,说明XG可促进细胞/基...
【英文摘要】
Objective:To test the effects of xyloglucan(XG),used as a new synthetic extracellular matrix(ECM)for adhesion of HepG2 cells and the albumin secretion of micro capsule HepG2 cells,ammonia elimination,cell proliferation and Cx32,E-cadherin gene expression of encapsulated HepG2 cells.Methods:HepG2 cells were seeded on different concentrations of XG coated surface and incubated for 4h,then the adherent cells were collected and quantified by BCA method for determining the adhesioin ratio.Micro capsules were pre...