PORCINE CARDIAC VALVULAR SUBENDOTHELIAL CELLS IN CULTURE - CELL ISOLATION AND GROWTH-CHARACTERISTICS
PORCINE CARDIAC VALVULAR SUBENDOTHELIAL CELLS IN CULTURE - CELL ISOLATION AND GROWTH-CHARACTERISTICS
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DOI：
10.1016/s0022-2828(87)80529-1
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发表时间：
1987-12-01
影响因子：
5
通讯作者：
HELGESON, SC
中科院分区：
文献类型：
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作者：
JOHNSON, CM;HANSON, MN;HELGESON, SC
我们从猪主动脉瓣的内皮下区域建立了培养系。这些细胞通过广泛的胶原酶消化瓣膜组织分离，并连续繁殖，具有稳定的形态。在稀疏培养中，瓣膜内皮下细胞类似于皮肤成纤维细胞。汇合时，瓣膜内皮下细胞形成类似血管平滑肌细胞的脊和桩。使用35 S-蛋氨酸的内源性体外标记实验表明，瓣膜内皮下细胞合成并释放了几种蛋白质，这些蛋白质在使用猪皮肤成纤维细胞和平滑肌细胞的平行实验中未观察到。使用猪主动脉瓣内皮细胞条件培养基的促分裂原试验表明，与皮肤成纤维细胞和平滑肌细胞相比，瓣膜内皮下细胞对体外瓣膜内皮细胞释放的生长因子特别敏感。瓣膜内皮下细胞对花生四烯酸的反应比皮肤成纤维细胞或平滑肌细胞释放的前列环素多10倍。我们的结论是，瓣膜内皮下细胞显示的功能，区分他们从其他培养的间充质细胞，这种培养系统将是有用的心脏瓣膜病的细胞基础的研究。
We established culture lines derived from the subendothelial region of the porcine aortic valve. These cells were isolated by extensive collagenase digestion of valvular tissue and were serially propagated with stable morphology. In sparse culture, valve subendothelial cells resembled skin fibroblasts. When confluent, the valve subendothelial cells formed ridges and piles similar to vascular smooth muscle cells. Endogenous in vitro labeling experiments using 35S-methionine showed that valve subendothelial cells synthesized and released several proteins not observed in parallel experiments using porcine skin fibroblasts and smooth muscle cells. Mitogen assays using media conditioned by porcine aortic valvular endothelial cells showed that the valve subendothelial cells, when compared to skin fibroblasts and smooth muscle cells, were particularly avid responders to the growth factors released by valve endothelial cells in vitro. The valve subendothelial cells also released 10-fold more prostacyclin in response to arachidonate than did skin fibroblasts or smooth muscle cells. We conclude that valve subendothelial cells show features that distinguish them from other cultured mesenchymal cells, and that this culture system will be useful for studies of the cellular basis of valvular heart disease.