Surowiec S M, Conklin B S, Li J S, Lin P H, Weiss V J, Lumsden A B, Chen C
Department of Surgery, Veterans Affairs Medical Center, Decatur, Georgia 30033, USA.
J Surg Res. 2000 Jan;88(1):34-41. doi: 10.1006/jsre.1999.5759.
Cell culture studies, ring studies, and indirect physiologic studies are the predominant models used to study human vascular tissue. Such studies are limited in their capacity to permit physiologic single-factor changes or to provide the proper mechanical stress or extracellular matrix present in normal tissues. We present a newly devised organ culture system that addresses these issues and permits survival of intact segments of human vascular tissue in a perfused environment. Our experience culturing human saphenous vein with this system is detailed.
Perfusion culture chambers were designed and constructed in our laboratory. Excess saphenous vein segments were collected from coronary artery bypass graft cases at our hospital and then mounted into our perfusion culture system for 0, 24, 48, 72, or 96 h. Vasomotor assays, hematoxylin and eosin staining, bromodeoxyuridine staining, and factor VIII staining were performed to assess tissue survival.
A total of 24 veins were cultured. Average vessel length was 5 cm. The vessels contracted and relaxed the following amounts: time 0 (6.7% contraction, 5.0% relaxation), 24 h (5.7%, 5.3%), 48 h (5.2%, 2.8%), 72 h (4.8%, 5.3%), 96 h (4.8%, 3.8%). Hematoxylin and eosin staining, bromodeoxyuridine staining, and factor VIII staining support the viability of the tissue segments.
A new perfusion organ culture system has been devised that permits survival of intact human venous tissue for periods up to 96 h. Studies that permit physiologic single-factor changes along with precise control of the hemodynamic environment are possible with this system.
细胞培养研究、环行研究和间接生理学研究是用于研究人体血管组织的主要模型。此类研究在允许生理学单因素变化或提供正常组织中存在的适当机械应力或细胞外基质方面能力有限。我们提出了一种新设计的器官培养系统,该系统解决了这些问题,并能使人体血管组织的完整节段在灌注环境中存活。我们使用该系统培养人隐静脉的经验在此详细介绍。
在我们实验室设计并构建了灌注培养室。从我院冠状动脉搭桥手术病例中收集多余的隐静脉节段,然后将其安装到我们的灌注培养系统中培养0、24、48、72或96小时。进行血管舒缩测定、苏木精-伊红染色、溴脱氧尿苷染色和因子VIII染色以评估组织存活情况。
共培养了24条静脉。平均血管长度为5厘米。血管收缩和舒张的幅度如下:0小时(收缩6.7%,舒张5.0%),24小时(5.7%,5.3%),48小时(5.2%,2.8%),72小时(4.8%,5.3%),96小时(4.8%,3.8%)。苏木精-伊红染色、溴脱氧尿苷染色和因子VIII染色均支持组织节段的存活能力。
已设计出一种新的灌注器官培养系统,该系统可使完整的人体静脉组织存活长达96小时。利用该系统可以进行允许生理学单因素变化并精确控制血流动力学环境的研究。