Nε-(carboxymethyl)lysine proliferated CD34+ cells from rat choroidal explant in culture
Nε-(carboxymethyl)lysine proliferated CD34+ cells from rat choroidal explant in culture
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DOI：
10.1248/bpb.27.1382
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发表时间：
2004-09-01
影响因子：
2
通讯作者：
Horiuchi, S
中科院分区：
文献类型：
--
作者：
Kobayashi, S;Shinohara, H;Horiuchi, S
本文观察了N-羧甲基赖氨酸-人血清白蛋白（CML-HSA）对体外培养的大鼠脉络膜血管新生的作用。将正常雄性Wistar大鼠脉络膜外植体培养在含有胎牛血清的Dulbecco改良Eagle培养基的纤维蛋白凝胶中，在存在或不存在CML-HSA的情况下。迁移的细胞在培养的第2天开始出芽，并以时间依赖性的方式从培养的脉络膜外植体中发育。从脉络膜外植体出芽和发育的细胞具有成纤维细胞的特征，其具有衰减的长胞质突起、长椭圆形核和许多膜结合的多形囊泡。用血管内皮细胞和内皮祖细胞的标志蛋白CD 34免疫组化染色纤维蛋白床中的减毒细胞未能显示阳性结果。而用胶原酶从纤维蛋白床分离的细胞用抗CD 34抗体特异性染色。培养3周后，分离的细胞在胶原凝胶上没有形成管状结构。CML-HSA可显著增加分离细胞总数和CD 34（+）细胞数以及血管样结构的数量。这些结果表明，CML-HSA从纤维蛋白凝胶中培养的脉络膜外植体过度产生未成熟的血管，其由CD 34（+）细胞组成。CML-HSA诱导的未成熟血管的形成可能与各种脉络膜疾病如年龄相关性黄斑变性有关。
Action of N-epsilon-(carboxymethyl)lysine-human serum albumin (CML-HSA) on neovascularization was investigated in cultured rat choroidal explant. Choroidal explants of normal male Wistar rats were cultured in fibrin gel with Dulbecco's modified Eagle medium containing fetal bovine serum in the presence or absence of CML-HSA. Migrated cells were budded from 2nd day in culture and developed from cultured choroidal explants in a time-dependent manner. Budded and developed cells from the choroidal explant had a feature of fibroblasts, which had attenuated long cytoplasmic processes, long ellipsoid nuclei and numerous membrane-bound polymorphic vesicles. Immunostaining of the attenuated cells in fibrin bed with CD34 (a marker protein of vascular endothelial cells and endothelial progenitor cells) failed to disclose positive result. However the cells which were isolated from fibrin bed by collagenase were specifically stained with anti-CD34 antibody. The isolated cells did not form tube-like structures on collagen gel by 3 weeks in culture. CML-HSA significantly increased the number of total isolated cells and CD34(+) cells as well as the number of vessel-like structures. These results indicate that CML-HSA overproduced immature blood vessels from cultured choroidal explants in fibrin gel, which consisted of CD34(+) cells. The CML-HSA-induced formation of immature blood vessel may be implicated in various choroidal diseases such as age-related macular degeneration.