|
- 2015
人淋巴管内皮细胞的分离和鉴定
|
Abstract:
摘要 目的:分离培养人淋巴管内皮细胞(LECs),为研究淋巴管新生在淋巴管瘤复发及肿瘤淋巴结转移中的作用奠定基础。方法:HE染色和免疫组织化学法了解人淋巴管瘤的组织病理特征;胶原酶消化法从淋巴管瘤组织中分离淋巴管内皮细胞;光镜和透射电镜观察淋巴管内皮细胞的形态和超微结构;MTT法检测细胞体外增殖。结果:HE染色显示舌淋巴管瘤中具有大量扩张的管腔,免疫组织化学染色显示管腔内皮细胞表达FLT-4、和LYVE-1;体外培养的淋巴管内皮细胞呈"铺路石"样外观;在透射电子显微镜下,观察到淋巴管内皮细胞特征性细胞器如weibel-palade小体、胞质小突和质膜小泡;MTT结果显示细胞于第3天开始进入增殖活跃期,第5天进入高峰期,第6、7天进入平台期;在Ⅰ型胶原凝胶中细胞形成管腔样结构。结论:应用胶原酶消化法可以有效的从淋巴管瘤组织中分离得到淋巴管内皮细胞
[1] | Garrafa E, Trainini L, Benetti A, Saba E, et al. Isolation, purification, and heterogeneity of human lymphatic endothelial cells from different tissues [J]. Lymphology, 2005, 38(4)∶159-166 |
[2] | Emirena G, Giulio A, Anna B, et al. Isolation and Characterization of Lymphatic Microvascular Endothelial Cells From Human Tonsils [J]. J Cell Physiol, 2006, 207(1)∶107-113 |
[3] | Chakravarti A, Bhargava R. Lymphangioma circumscriptum of the tongue in children: successful treatment using intralesional bleomycin [J]. Int J Pediatr Otorhinolaryngol, 2013, 77(8)∶1367-1369 |
[4] | Faul JL, Berry GJ, Colby TV, et al. Thoracic Lymphangiomas, Lymphangiectasis, Lymphangiomatosis, and Lymphatic Dysplasia Syndrome [J]. Am J Respir Crit Care Med, 2000, 161(3)∶1037-1046 |
[5] | Simona P, Pascal B, Paula V, et al. Molecular characterization of lymphatic endothelial cells [J]. PNAS, 2002, 99(25)∶16069-16074 |
[6] | Ernst K, Silvana BG, Marion G, et al. Isolation and Characterization of Dermal Lymphatic and Blood Endothelial Cells Reveal Stable and Functionally Specialized Cell Lineages [J]. J. Exp. Med, 2001, 194(6)∶797-808 |
[7] | Johnson MG, Walker MA Lymphatic endothelial and smooth muscle cells in tissue culture [J]. In Vitro, 1984, 20(7)∶566-562 |
[8] | Leak, LV. The transport of exogenous peroxidase across the blood-tissuelymph interface [J]. J. Ultrastruct. Res, 1972, 39(1)∶24-42 |
[9] | Sironi M, Conti A, Bernasconi S, Fra AM, et al. Generation and characterization of a mouse lymphatic endothelial cell line [J]. Cell Tissue Res, 2006, 325(1)∶91-100 |
[10] | Mitsukawa N, Satoh K. New treatment for cystic lymphangiomas of the face and neck: cyst wall rupture and cyst aspiration combined with sclerotherapy [J]. J Craniofac Surg, 2012, 23(4)∶1117-1119 |
[11] | Makinen T, Veikkola T, Mustjoki S et al. Isolated lymphatic endothelial cells transduce growth, survival and migratory signals via the VEGF-C/D receptor VEGFR-3 [J]. EMBO J, 2001, 20(17)∶4762-4773 |
[12] | Thornton SC,Mueller SN and Levine EM. Human endothelial cells. Use of heParin in cloning and long-term serial cultivation [J]. Science, 1983, 222(4624)∶623-625 |
[13] | Jones BE, Yong, LC. Culture and characterization of bovine mesenteric lymphatic endothelium [J]. In vitro cellular dev boil, 1987, 23(10)∶698-706 |
[14] | Djoneidi M, Brodt P. Isolation and characterization of rat lymphatic endothelial cells [J]. Microcirc Endothelium Lymphatics, 1991, 7(4-6)∶161-182 |