Bernik M B, Kwaan H C
J Clin Invest. 1969 Sep;48(9):1740-53. doi: 10.1172/JCI106140.
Human tissues and cells from pre- and postnatal life were cultivated and studied for plasminogen activator activity. Cultures were obtained from kidney, renal blood vessels, ureter, bladder, lung, and heart. Local activator activity of cells was demonstrated by histochemical techniques. Activator released by cells into the supernatant culture media was assayed by fibrin plate techniques and was investigated for immunological identity using specific antisera to an activator of human origin, urokinase (UK). Plasminogen activator was produced in primary cultures where cells retain specific functions and generally reflect the enzyme pattern of the tissues of origin. Cells from fetal and adult sources were found to yield activator antigenically identical to UK, as well as activator activity which differed from that of UK in immunoassays and which may represent tissue type activator. Such activity was released after injury or death of cells while UK was produced in cultures containing live, metabolizing cells. Primary cultures of kidney confirmed that this organ is a rich source of UK and demonstrated, in addition, that UK is produced from the early stages of gestation and in increasing amounts thereafter. However, primary cultures also demonstrated that the ability to produce UK is not limited to the kidney but is a function of cells which are distributed widely in body tissues. Thus, activator antigenically identical to UK accumulated progressively after many refeedings in culture supernates of fetal lung and ureter, as well as in supernates of renal blood vessels of adults. These findings indicate continuous formation of UK by the cultured cells and, furthermore, provide evidence of UK production in blood vessels. In cultures from other tissues, particularly those from fetal heart and adult lung and bladder, investigation of activator was hindered by inhibitory activity which accumulated in the supernates. Such activity was derived from cells in culture and was directed selectively against UK, indicating that inhibitor as well as UK are produced by cells of various organs of the body. Plasminogen activator also was produced by serially propagated cells, diploid and heteroploid. However, only diploid cell lines retained activator activity of the original tissues and continued to produce activator antigenically identical to UK. In contrast, heteroploid line appeared to have lost the ability to form UK by yielded activator activity that differed from that of UK in immunoassays. Serially propagated cells thus provide an additional tool for in vitro study of plasminogen activator and may facilitate investigation of the fibrinolytic system in man.
对来自产前和产后生活的人体组织和细胞进行培养，并研究其纤溶酶原激活物活性。培养物取自肾脏、肾血管、输尿管、膀胱、肺和心脏。通过组织化学技术证明细胞的局部激活物活性。用纤维蛋白平板技术测定细胞释放到上清培养基中的激活物，并使用针对人源激活物尿激酶（UK）的特异性抗血清研究其免疫学特性。纤溶酶原激活物在原代培养物中产生，其中细胞保留特定功能，并通常反映起源组织的酶模式。发现来自胎儿和成人来源的细胞产生与UK抗原相同的激活物，以及在免疫测定中与UK不同的激活物活性，这可能代表组织型激活物。这种活性在细胞损伤或死亡后释放，而UK是在含有活的、代谢活跃细胞的培养物中产生的。肾脏的原代培养证实该器官是UK的丰富来源，此外还表明UK从妊娠早期开始产生，此后产量不断增加。然而，原代培养也表明产生UK的能力不限于肾脏，而是广泛分布于身体组织中的细胞的功能。因此，与UK抗原相同的激活物在胎儿肺和输尿管的培养上清液以及成人肾血管的上清液中经过多次再喂养后逐渐积累。这些发现表明培养细胞持续形成UK，此外，还提供了血管中产生UK的证据。在来自其他组织的培养物中，特别是来自胎儿心脏、成人肺和膀胱的培养物中，上清液中积累的抑制活性阻碍了对激活物的研究。这种活性源自培养中的细胞，并且选择性地针对UK，表明抑制剂以及UK都是由身体各个器官的细胞产生的。纤溶酶原激活物也由连续传代的细胞（二倍体和异倍体）产生。然而，只有二倍体细胞系保留了原始组织的激活物活性，并继续产生与UK抗原相同的激活物。相比之下，异倍体细胞系似乎失去了形成UK的能力，其产生的激活物活性在免疫测定中与UK不同。因此，连续传代的细胞为纤溶酶原激活物的体外研究提供了额外的工具，并可能有助于对人体纤溶系统的研究。
