Kitaguchi H, Funahara Y, Butthep P, Bunyaratvej A
Department of Physiology 1, Kobe University School of Medicine, Japan.
Southeast Asian J Trop Med Public Health. 1992;23 Suppl 2:95-100.
We cultivated endothelial cells of human umbilical vein origin in the presence of red blood cells and platelet-rich plasma, and observed the phenomena which occurred in the petri dish under phase contrast microscopy. Many small particles were observed after an overnight incubation. We washed the dish two times, then added thrombin to the dish. The network of thread-like strands appeared within 10 to 20 minutes of the addition, and at the same time the small particles adhered on the surface of the strands, swelled and fused gradually to cover the surface of the strands completely. Within 30 to 60 minutes the network of the strands changed into a capillary-like structure. These phenomena were not observed if we omitted red blood cells or platelet-rich plasma. Studies by transmission electron microscopy revealed that the inner surface of the lumen of the structure was covered with cells. The cells isolated from the lumen by trypsin grew to confluence in the conventional culture medium, and showed vWF antigen on their surface. These observations indicated that the method described is useful for in vitro study of angiogenesis.
我们在红细胞和富含血小板血浆存在的情况下培养人脐静脉来源的内皮细胞,并在相差显微镜下观察培养皿中发生的现象。过夜孵育后观察到许多小颗粒。我们将培养皿洗涤两次,然后向培养皿中加入凝血酶。加入凝血酶后10至20分钟内出现丝状网络,同时小颗粒附着在丝的表面,逐渐肿胀并融合以完全覆盖丝的表面。30至60分钟内,丝的网络变成毛细血管样结构。如果我们省略红细胞或富含血小板血浆,则不会观察到这些现象。透射电子显微镜研究表明,该结构管腔的内表面覆盖有细胞。通过胰蛋白酶从管腔中分离出的细胞在传统培养基中生长至汇合,并在其表面显示vWF抗原。这些观察结果表明,所述方法对于血管生成的体外研究是有用的。