Booyse F M, Quarfoot A J, Bell S, Fass D N, Lewis J C, Mann K G, Bowie E J
Proc Natl Acad Sci U S A. 1977 Dec;74(12):5702-6. doi: 10.1073/pnas.74.12.5702.
Aortic endothelial cells from normal pigs and pigs with von Willebrand disease have been established in long-term cultures. Both cultures appeared similar in terms of general growth characteristics, morphologic features and ultrastructure. Immunofluorescent staining of these cultures with chicken (or rabbit) antiporcine ristocetin-Willebrand factor sera (or IgG) resulted in extensive perinuclear staining of the cells in both cultures. Additionally, staining of semiconfluent cultures of normal cells for ristocetin-Willebrand factor revealed an extensive meshwork of distinct, immunologically identifiable ristocetin-Willebrand factor-containing filaments between cells. Immunoreactive material was considerably decreased and more diffuse between cells in semiconfluent cultures from affected pigs. Through immunocytochemical staining with peroxidase-coupled antiserum, the filaments (of indeterminate length) were found to have a diameter of approximately 300 A. Finally, washed porcine platelets interacted extensively with scrape-damaged cultures of affected endothelial cells. This interaction of platelets with damaged normal cultures was abolished by pretreatment of the cultures with rabbit antiporcine ristocetin-Willebrand factor IgG.
已成功建立了来自正常猪和患有血管性血友病的猪的主动脉内皮细胞的长期培养体系。两种培养体系在总体生长特性、形态特征和超微结构方面表现相似。用鸡（或兔）抗猪瑞斯托霉素-血管性血友病因子血清（或IgG）对这些培养物进行免疫荧光染色，结果两种培养物中的细胞均出现广泛的核周染色。此外，对正常细胞的亚汇合培养物进行瑞斯托霉素-血管性血友病因子染色，结果显示细胞间存在广泛的、由不同的、可通过免疫鉴定的含瑞斯托霉素-血管性血友病因子的细丝组成的网络。在患病猪的亚汇合培养物中，细胞间的免疫反应性物质显著减少且更加弥散。通过用与过氧化物酶偶联的抗血清进行免疫细胞化学染色，发现这些细丝（长度不确定）的直径约为300埃。最后，洗涤后的猪血小板与受损的患病内皮细胞刮擦损伤培养物广泛相互作用。用兔抗猪瑞斯托霉素-血管性血友病因子IgG预处理培养物后，血小板与受损正常培养物之间的这种相互作用被消除。
