Valdes-Camin R, Callahan J B, Ebert R F
Organon Teknika/Biotechnology Research Institute, Rockville, MD 20850.
Blood Coagul Fibrinolysis. 1994 Aug;5(4):617-24.
High-sensitivity thromboplastin reagents suitable for use in the prothrombin time (PT) assay are typically prepared from human brain and placenta, tissues that are in limited supply and subject to viral contamination. Cloning and expression of recombinant human tissue factor (TF) has enabled production of a new generation of thromboplastin reagents whose performance and utility are under active investigation. The purpose of this study was to determine the feasibility of producing a sensitive human thromboplastin reagent from a non-recombinant source: cultured human cells. Several human cell lines with apparently high constitutive TF synthesis were identified, and a viable thromboplastin reagent (Humaplastin) was produced from a human lung cell line via a non-conventional process that did not require reconstitution or rehydration of TF in cell membranes. When calibrated against BCT/253, a human brain international reference thromboplastin, Humaplastin exhibited a mean normal prothrombin time of 12.6 +/- 0.7 s (mean +/- SD: n = 20) and an International Sensitivity Index of 1.09 +/- 0.019. The performance of this reagent was well correlated (r = 0.983) with Thromborel S, a commercially available human placental thromboplastin reagent. Orthogonal least squares regression of the log PT values from the placental thromboplastin reagent versus Humaplastin and two recombinant TF-based thromboplastin reagents suggested that the latter three reagents are somewhat more sensitive than the placental thromboplastin reagent, although such differences should not be expected to have a significant impact on clinical utility. It is concluded that cultured human lung cells represent a suitable source of tissue thromboplastin for production of a high-sensitivity non-recombinant thromboplastin reagent.
适用于凝血酶原时间（PT）测定的高灵敏度凝血活酶试剂通常由人脑和胎盘制备，而这些组织供应有限且易受病毒污染。重组人组织因子（TF）的克隆和表达使得新一代凝血活酶试剂得以生产，其性能和效用正在积极研究中。本研究的目的是确定从非重组来源——培养的人细胞生产敏感的人凝血活酶试剂的可行性。鉴定出几种明显具有高组成性TF合成的人细胞系，并通过一种非常规方法从人肺细胞系生产出一种可行的凝血活酶试剂（Humaplastin），该方法不需要对细胞膜中的TF进行重构或复水。当以人脑国际参考凝血活酶BCT/253进行校准时，Humaplastin的平均正常凝血酶原时间为12.6±0.7秒（平均值±标准差：n = 20），国际灵敏度指数为1.09±0.019。该试剂的性能与市售人胎盘凝血活酶试剂Thromborel S具有良好的相关性（r = 0.983）。对胎盘凝血活酶试剂与Humaplastin以及两种基于重组TF的凝血活酶试剂的log PT值进行正交最小二乘回归分析表明，后三种试剂比胎盘凝血活酶试剂稍敏感，尽管预计这种差异对临床效用不会有显著影响。结论是，培养的人肺细胞是生产高灵敏度非重组凝血活酶试剂的合适组织凝血活酶来源。
