Castronuovo John J, Smith Thomas J, Price Ray M
Department of Surgery, Reeves Laboratory for Surgical Research, Morristown Memorial Hospital, 100 Madison Ave., Morristown, NJ 07962-1956, USA.
J Vasc Surg. 2002 Jan;35(1):152-7.
The purpose of this study was the validation of the physiologic appropriateness of in vitro organ culture of human saphenous vein as a model with the demonstration of the occurrence of the processes of cell proliferation, remodeling, and hyperplasia.
Saphenous vein from 28 patients was cross-sectioned into seven 2-mm segments and maintained in organ culture for 2 days or 2 weeks. Three organ culture media were used: a chemically well-defined medium (RPMI-1640) and the same medium supplemented with the undefined protein-containing supplements fetal bovine serum (FBS) or pooled adult human plasma (type AB). The outcome measures at 2 days and 2 weeks were compared with measurements of segments from the same vein at the time of harvest. Excess saphenous vein harvested for arterial bypass grafting was obtained after approval of the study protocol by the Institutional Review Board. Cell proliferation was measured with immunostaining for proliferating cell nuclear antigen. Remodeling and intimal hyperplasia were measured with micromorphometric comparisons of vein segment cross-sectional area before and after organ culture.
There was no evidence of cell death or tissue degeneration on histologic examination of the cultured vein segments. Cell proliferation, expressed as proliferation index (PI; positive proliferating cell nuclear antigen nuclei/total nuclei), significantly increased as compared with freshly harvested vein after 2 days of culture in undefined, protein-supplemented media (mean PI, 42.4 +/- 7.4%; P <.001). A significant increase in cell proliferation did not occur in the defined, unsupplemented medium until 2 weeks (mean PI, 16.2 +/- 7.1%; P <.001). The cross-sectional area of the vein wall increased during culture in all media. A statistically significant increase in the cross-sectional area of the vein wall occurred during culture with plasma (P <.001) and FBS supplementation (P =.002). The increase in the cross-sectional area of the vein in defined media was almost statistically significant (P =.089). A significant increase was seen in the cross-sectional area of the media (P =.006) and adventitia (P =.030) of veins cultured with plasma supplementation and in the cross-sectional area of the adventitia (P =.034) of veins cultured with FBS supplementation.
These results show that human saphenous vein in culture is viable, shows cell proliferation, and exhibits remodeling of the layers of the vein wall. This is the first report to document hyperplasia in human vascular tissue cultured in a defined medium.
本研究的目的是验证人隐静脉体外器官培养作为一种模型在生理上的适宜性，并证明细胞增殖、重塑和增生过程的发生。
将28例患者的隐静脉横切成7段2毫米长的片段，并在器官培养中维持2天或2周。使用了三种器官培养基：一种化学成分明确的培养基（RPMI-1640）以及添加了不明确含蛋白质补充剂的相同培养基，即胎牛血清（FBS）或混合成人血浆（AB型）。将2天和2周时的结果测量值与收获时同一静脉片段的测量值进行比较。在研究方案经机构审查委员会批准后，获取用于动脉搭桥移植术多余的隐静脉。通过增殖细胞核抗原免疫染色测量细胞增殖。通过对器官培养前后静脉片段横截面积的微观形态学比较来测量重塑和内膜增生。
对培养的静脉片段进行组织学检查，未发现细胞死亡或组织退化的证据。在添加了不明确蛋白质补充剂的培养基中培养2天后，以增殖指数（PI；增殖细胞核抗原阳性细胞核数/总细胞核数）表示的细胞增殖与新鲜收获的静脉相比显著增加（平均PI，42.4±7.4%；P<.001）。在未添加补充剂的明确培养基中，直到2周时细胞增殖才显著增加（平均PI, 16.2±7.1%；P<.001）。在所有培养基中培养期间，静脉壁的横截面积均增加。在添加血浆（P<.001）和FBS补充剂（P =.002）的培养过程中，静脉壁的横截面积有统计学显著增加。在添加明确培养基中，静脉横截面积的增加几乎具有统计学显著性（P =.089）。在添加血浆培养的静脉中膜（P =.006）和外膜（P =.030）的横截面积以及添加FBS培养的静脉外膜（P =.034）的横截面积有显著增加。
这些结果表明，培养中的人隐静脉是有活力的，显示出细胞增殖，并表现出静脉壁各层的重塑。这是第一份记录在明确培养基中培养的人血管组织增生的报告。
