HUVEC or human artery endothelial cells (HUAECs) and human brain-deriv的简体中文翻译

HUVEC or human artery endothelial c

HUVEC or human artery endothelial cells (HUAECs) and human brain-derived vascular pericytes or bovine retinal pericytes are trypsinized and seeded within 3D collagen matrices at a density of 2 × 106 cells/ml for EC and 4 × 105 cells/ml for pericytes. Both types of EC or pericytes work very well in this assay system. FGF-2 and stromal-derived factor-1α (SDF-1α) are each added at 200 ng/ml into the collagen gel mix (2.5 mg/ml of rat collagen type I). Twenty-five μl of gel is added into each individual 96 well (A/2 plates from Costar). After 30 min of equilibration in a CO2 incubator, culture medium is added (100 μl per well) which is Medium 199 (100 μl per well supplemented with 1:250 dilution of Reduced Serum Supplement II) [160], and 40 ng/ml each of FGF-2, stem cell factor (SCF), interleukin-3 (IL-3), and 50 μg/ml of ascorbic acid are added. Cultures are then placed in a CO2 incubator for 72–120 h. After this time, cultures are fixed with 3% paraformaldehyde in PBS (120 μl per well). Cultures can then be imaged under fluorescence to quantify EC tube area and pericyte recruitment under the various conditions of treatment. Individual cultures can be immunostained with various antibodies directed to EC or pericyte cell surfaces and/or ECM antigens (i.e., basement membrane proteins such as fibronectin, laminin, and collagen type IV) [153]. Alternatively, cultures can be used to make lysates using SDS sample buffer or used to isolate total RNA to perform gene expression experiments [153]. Real-time movies can be performed to examine pericyte recruitment and measure pericyte motility during capillary assembly and maturation [154]. In the latter case, nuclear GFP-labeled pericytes can be utilized to track pericyte motility
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结果 (简体中文) 1: [复制]
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<br>用胰蛋白酶消化HUVEC或人动脉内皮细胞(HUAEC)以及人脑衍生的血管周细胞或牛视网膜周细胞,并以<br>2×106 <br>细胞/ ml 的密度接种到3D胶原蛋白基质中,并以4×105 <br>细胞/ ml 的密度接种到周细胞中。两种类型的EC <br>或周细胞在该测定系统中均能很好地发挥作用。将FGF-2和基质衍生因子1α <br>(SDF-1α)分别以200 ng / ml的浓度添加到胶原蛋白凝胶混合物(2.5 mg / ml的<br>I型大鼠胶原蛋白)中。将25μl的凝胶添加到每个单独的96孔中(来自Costar的A / 2板)。<br>在CO2培养箱中平衡30分钟后,添加培养基(每孔100μl)<br>,即培养基199(每孔100μl,补充1:250稀释的还原血清)<br>补品II)[160],分别添加40 ng / ml FGF-2,干细胞因子(SCF),白介素3 <br>(IL-3)和50μg/ ml抗坏血酸。然后将培养物置于二氧化碳培养箱<br>中72–120小时。此时间之后,将培养物用PBS中的3%低聚甲醛固定(<br>每孔120μl )。然后可以在荧光下使培养物成像,以量化<br>在各种治疗条件下的EC管面积和周细胞募集。个别培养物可用<br>针对EC或周细胞表面和/或ECM <br>抗原(即,诸如纤连蛋白,层粘连蛋白和<br>IV 型胶原的基底膜蛋白)的各种抗体免疫染色[153]。或者,可以使用SDS样品缓冲液或<br>用于分离总RNA进行基因表达实验[153]。<br>可以执行实时电影检查<br>毛细血管组装和成熟过程中的周细胞募集并测量周细胞运动性[154]。在后一种情况下,核GFP标记的周细胞<br>可用于追踪周细胞运动
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结果 (简体中文) 2:[复制]
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HUVEC or human artery endothelial cells (HUAECs) and human brain-derived vascular <br>pericytes or bovine retinal pericytes are trypsinized and seeded within 3D collagen matrices <br>at a density of 2 × 106<br> cells/ml for EC and 4 × 105<br> cells/ml for pericytes. Both types of EC <br>or pericytes work very well in this assay system. FGF-2 and stromal-derived factor-1α <br>(SDF-1α) are each added at 200 ng/ml into the collagen gel mix (2.5 mg/ml of rat collagen <br>type I). Twenty-five μl of gel is added into each individual 96 well (A/2 plates from Costar). <br>After 30 min of equilibration in a CO2 incubator, culture medium is added (100 μl per well) <br>which is Medium 199 (100 μl per well supplemented with 1:250 dilution of Reduced Serum <br>Supplement II) [160], and 40 ng/ml each of FGF-2, stem cell factor (SCF), interleukin-3 <br>(IL-3), and 50 μg/ml of ascorbic acid are added. Cultures are then placed in a CO2 incubator <br>for 72–120 h. After this time, cultures are fixed with 3% paraformaldehyde in PBS (120 μl <br>per well). Cultures can then be imaged under fluorescence to quantify EC tube area and <br>pericyte recruitment under the various conditions of treatment. Individual cultures can be <br>immunostained with various antibodies directed to EC or pericyte cell surfaces and/or ECM <br>antigens (i.e., basement membrane proteins such as fibronectin, laminin, and collagen type <br>IV) [153]. Alternatively, cultures can be used to make lysates using SDS sample buffer or <br>used to isolate total RNA to perform gene expression experiments [153]. Real-time movies <br>can be performed to examine pericyte recruitment and measure pericyte motility during <br>capillary assembly and maturation [154]. In the latter case, nuclear GFP-labeled pericytes <br>can be utilized to track pericyte motility
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结果 (简体中文) 3:[复制]
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人血管内皮细胞(HUVEC)和人脑血管<br>周细胞或牛视网膜周细胞被胰蛋白酶化并植入3D胶原基质中<br>密度为2×106时<br>EC和4×105细胞/ml<br>周细胞/ml。两种类型的EC<br>或者周细胞在这个分析系统中工作得很好。FGF-2与基质衍生因子-1α<br>(SDF-1α)分别以200 ng/ml加入胶原凝胶混合物(2.5 mg/ml大鼠胶原)中<br>类型I)。在每个96孔中加入25μl凝胶(A/2板来自Costar)。<br>在二氧化碳培养箱中平衡30分钟后,添加培养基(100μl/孔)<br>即培养基199(每孔100μl,补充1:250稀释的还原血清<br>补充II)[160]和40 ng/ml的FGF-2、干细胞因子(SCF)、白细胞介素-3<br>(IL-3)和50μg/ml抗坏血酸。然后将培养物放在二氧化碳培养箱中<br>72-120小时。在此之后,用3%多聚甲醛在PBS(120μl)中固定培养物<br>每口井)。培养物可在荧光下成像,以量化EC管面积和<br>不同治疗条件下周细胞的募集。个人文化可以是<br>用针对EC或周细胞表面和/或ECM的各种抗体进行免疫染色<br>抗原(即基底膜蛋白,如纤维连接蛋白、层粘连蛋白和胶原类型<br>四) [153]。或者,培养物可用于使用SDS样品缓冲液或<br>用于分离总RNA进行基因表达实验[153]。实时电影<br>可用于检测周细胞募集和检测<br>毛细管组装和成熟[154]。在后一种情况下,核GFP标记周细胞<br>可用于跟踪周细胞运动<br>
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