Remy M, Bordenave L, Bareille R, Lauroua C, Lefebvre F, Guerin V, Baquey C
INSERM-U. 306, Université de Bordeaux II, France.
Biomaterials. 1995 Feb;16(3):193-9. doi: 10.1016/0142-9612(95)92117-o.
In the field of in vitro biocompatibility testing, the investigation of cell response at the interface with a biomaterial is of great importance; there is a need for standard conditions and thus of well-defined and reliable sources of materials for an objective evaluation of cellular function. Thrombin is often used in vitro as a stimulating agent to check the specific functions of cultured endothelial cells. In the present work, and in order to select a thrombin of commercial origin, two criteria were borne in mind: purity towards the presence of the von Willebrand factor (vWF) and effectiveness towards vWF release by human umbilical venous endothelial cells (HUVEC) that have been submitted to four human commercial thrombins. We detected the presence of vWF in some thrombin solutions that have not yet been in contact with HUVEC. The different thrombins contained vWF antigen ranging from less than 0.1 mUnit per NIH unit of thrombin (from Diagnostica Stago and Sigma Chemical Co.) to 10-20 mUnit per NIH unit of Fibrindex thrombin (from Ortho Diagnostic Systems). Thus, if vWF is present in commercial thrombins, it contributes to and overestimates the vWF appearance in the media resulting from cell stimulation. Consequently, we fixed on thrombin from Diagnostica Stago for further studies involving HUVEC on biomaterials.
在体外生物相容性测试领域，研究生物材料界面处的细胞反应非常重要；为了客观评估细胞功能，需要标准条件以及定义明确且可靠的材料来源。凝血酶在体外常被用作刺激剂，以检查培养的内皮细胞的特定功能。在本研究中，为了选择市售来源的凝血酶，考虑了两个标准：针对血管性血友病因子（vWF）存在的纯度，以及对已接触四种市售人凝血酶的人脐静脉内皮细胞（HUVEC）释放vWF的有效性。我们在一些尚未与HUVEC接触的凝血酶溶液中检测到了vWF的存在。不同的凝血酶所含vWF抗原量从每NIH单位凝血酶低于0.1 mUnit（来自Diagnostica Stago和Sigma Chemical Co.）到每NIH单位Fibrindex凝血酶10 - 20 mUnit（来自Ortho Diagnostic Systems）不等。因此，如果市售凝血酶中存在vWF，它会导致并高估细胞刺激后培养基中vWF的出现量。因此，我们选定Diagnostica Stago的凝血酶用于进一步涉及HUVEC与生物材料的研究。
