BASAL TISSUE FACTOR EXPRESSION IN ENDOTHELIAL-CELL CULTURES IS CAUSED BY CONTAMINATING SMOOTH-MUSCLE CELLS - REDUCTION BY USING CHYMOTRYPSIN INSTEAD OF COLLAGENASE
BASAL TISSUE FACTOR EXPRESSION IN ENDOTHELIAL-CELL CULTURES IS CAUSED BY CONTAMINATING SMOOTH-MUSCLE CELLS - REDUCTION BY USING CHYMOTRYPSIN INSTEAD OF COLLAGENASE
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DOI：
10.1016/0049-3848(95)00192-t
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发表时间：
1995-12-01
影响因子：
7.5
通讯作者：
BOM, VJJ
中科院分区：
文献类型：
--
作者：
MULDER, AB;BLOM, NR;BOM, VJJ
在内皮细胞培养中发现的基底组织因子（TF）表达与在体内未受干扰的内皮细胞中检测不到TF之间存在差异。我们证明了内皮细胞培养中基底TF的表达起源于污染细胞。这些细胞经超微结构和流式细胞术鉴定为平滑肌细胞。从胶原酶处理的人脐带血管中获得细胞培养物。组织学研究显示，胶原酶处理后，基底膜被消化，静脉某些区域的基础结构被破坏。我们选择胰凝乳酶作为分离内皮细胞的替代方法。使用胰凝乳蛋白酶，内皮细胞选择性地消失，基底膜不受干扰。此外，使用凝乳胰蛋白酶代替胶原酶使基础TF活性水平降至最低。综上所述，我们证明了内皮细胞培养中基底TF表达是由污染平滑肌细胞引起的。这种污染可以强烈地减少使用凝乳胰蛋白酶代替胶原酶分离内皮细胞。
A discrepancy exists between basal tissue factor (TF) expression found in endothelial cell cultures and the failure to detect TF in unpertubated endothelial cells in vivo. We demonstrated that basal TF expression in endothelial cell cultures originated from contaminating cells. These cells were ultrastructurally and flowcytometrically identified as smooth muscle cells. The cell cultures had been obtained from collagenase-treated human umbilical cord vessels. Histologic studies revealed that after collagenase treatment the basement membrane was digested and underlying structures were disrupted at some areas of the vein. We selected chymotrypsin as an alternative for the isolation of endothelial cells. Using chymotrypsin, the endothelial lining was selectively lost leaving the basement membrane undisturbed. Furthermore, use of chymotrypsin instead of collagenase minimized the level of basal TF activity. Taken together, we demonstrated that basal TF expression in endothelial cell cultures is caused by contaminating smooth muscle cells. This contamination can strongly be reduced using chymotrypsin instead of collagenase for isolation of endothelial cells.