Development and characterization of endothelial cells from rat microlymphatics.
Development and characterization of endothelial cells from rat microlymphatics.
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DOI：
10.1089/153968503321642606
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发表时间：
2003-01-01
影响因子：
1.4
通讯作者：
Zawieja, D
中科院分区：
文献类型：
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作者：
Hayes, H;Kossmann, E;Zawieja, D
背景：淋巴内皮对于淋巴系统的功能很重要，包括淋巴重塑、血管张力控制以及液体、大分子和细胞的淋巴运动。许多这些事件主要发生在微淋巴系统的水平。为了评估微淋巴内皮的作用，合适的培养细胞系将是有用的。我们开发了一种从直径约 100 微米的微观淋巴管中分离和培养内皮细胞的技术。方法和结果：为了分离大鼠肠系膜淋巴内皮细胞 (RMLEC)，将大鼠麻醉并小心地将肠系膜取出。找到合适的微淋巴管，并从周围的肠系膜上仔细地进行显微解剖。将容器仔细清洗、插管、翻转，然后在涂有明胶的塑料培养皿上孵育，直到小片细胞从容器中迁移出来（3-4天后）。然后取出外植的容器。培养剩余的细胞并筛选内皮表型。非内皮细胞被破坏。其余细胞的内皮性质通过以下方式验证：1) 形态，2) 荧光乙酰化 LDL 的摄取，3) von Wille-brand 因子、PECAM-1、ecNOS、LYVE-1、VEGFR-3 染色，以及 4) 基本上阴性的 α-血管平滑肌肌动蛋白染色。通过定义的 RMLEC，然后使用 PCR 和免疫荧光测定 RMLEC 上存在的粘附分子的概况。结论：我们开发并部分表征了培养的微淋巴内皮细胞系。 RMLEC 表达已知的内皮和淋巴特异性标记物以及以下粘附分子：N-钙粘蛋白、E-钙粘蛋白、PECAM-1、α-连环蛋白、β-连环蛋白、γ-连环蛋白、p120 和各种整合素。
BACKGROUND: The lymphatic endothelium is important to the functioning of the lymphatic system, including lymphatic remodeling, control of vessel tone, and lymphatic movement of fluids, macromolecules, and cells. Many of these events occur principally at the level of the microlymphatics. To evaluate the role of the microlymphatic endothelium, a suitable cultured cell line would be useful. We have developed a technique to isolate and culture endothelial cells from microscopic lymphatics, approximately 100 microm in diameter.METHODS AND RESULTS: To isolate the rat mesenteric lymphatic endothelial cells (RMLEC), the rat was anesthetized and the mesentery carefully exteriorized. A suitable microlymphatic was located and carefully microdissected from the surrounding mesentery. The vessel was carefully cleaned, cannulated, everted, and then incubated on a gelatin-coated plastic culture dish until small patches of cells migrated off of the vessel (3-4 days later.) The explanted vessel was then removed. The remaining cells were cultured and screened for endothelial phenotype. Nonendothelial cells were destroyed. The endothelial nature of the remaining cells was verified by: 1) morphology, 2) uptake of fluorescent acetylated-LDL, 3) staining for von Wille-brand factor, PECAM-1, ecNOS, LYVE-1, VEGFR-3, and 4) essentially negative alpha-vascular smooth muscle actin staining. The defined RMLEC were passed and the profile of adhesion molecules present on the RMLEC was then determined using PCR and immunofluorescence.CONCLUSIONS: We developed and partially characterized a line of cultured microlymphatic endothelium. RMLEC express known endothelial- and lymphatic-specific markers as well as the following adhesion molecules: N-cadherin, E-cadherin, PECAM-1, alpha-catenin, beta-catenin, gamma-catenin, p120, and a variety of integrins.