Baxter Healthcare Corporation, BioSurgery Division, Westlake Village, California, USA.
Tissue Eng Part A. 2011 Feb;17(3-4):429-37. doi: 10.1089/ten.TEA.2010.0113. Epub 2011 Jan 13.
Over the last several years, our in vitro and in vivo studies have focused on optimizing the use of fibrin to deliver cells. We have shown that some three-dimensional (3D) fibrin constructs with specific fibrinogen and thrombin concentration support robust proliferation of normal human dermal fibroblasts, whereas different fibrinogen and thrombin concentrations support high mesenchymal stem cell proliferation in 3D fibrin constructs. In this article, we found that normal human epithelial keratinocytes proliferate well in 3D fibrin constructs consist of fibrinogen concentration ranging from 17 to 33 mg/mL and thrombin concentration of 1 U/mL. Further, using a new proliferation assay, we studied the proliferation of fibroblasts and keratinocytes cocultured in various 3D fibrin constructs of different fibrinogen and thrombin concentrations. We found that 3D fibrin constructs with a range of fibrinogen concentration (5-34 mg/mL) and a thrombin concentration of 1 U/mL produce an optimal cell proliferation for both cell types when cocultured. This profile of proliferation is different from that seen when keratinocytes or fibroblasts are incorporated separately in 3D fibrin constructs. In conclusion, we found that one needs to choose the fibrinogen and thrombin concentration carefully depending on the cell type to deliver; that is, different fibrin constructs with different fibrinogen and thrombin concentration are required to deliver fibroblasts or keratinocytes alone or to codeliver both cell types. Moreover, there seems to be a cross-talk between keratinocytes and fibroblasts when they are cointroduced in 3D fibrin constructs. This feedback could be due to the effects of growth factors produced by the two cell types in the 3D fibrin constructs.
在过去的几年中，我们的体外和体内研究集中在优化纤维蛋白的使用以递送细胞。我们已经表明，一些具有特定纤维蛋白原和凝血酶浓度的三维（3D）纤维蛋白构建体支持正常人类真皮成纤维细胞的旺盛增殖，而不同的纤维蛋白原和凝血酶浓度支持 3D 纤维蛋白构建体中的高间充质干细胞增殖。在本文中，我们发现正常的人类上皮角质形成细胞在纤维蛋白原浓度范围为 17 至 33mg/mL 和凝血酶浓度为 1U/mL 的 3D 纤维蛋白构建体中增殖良好。此外，使用新的增殖测定法，我们研究了在不同纤维蛋白原和凝血酶浓度的各种 3D 纤维蛋白构建体中培养的成纤维细胞和角质形成细胞的共培养增殖。我们发现，纤维蛋白原浓度范围为 5-34mg/mL 和凝血酶浓度为 1U/mL 的 3D 纤维蛋白构建体在共培养时为两种细胞类型产生最佳的细胞增殖。这种增殖模式与单独将角质形成细胞或成纤维细胞掺入 3D 纤维蛋白构建体中所观察到的模式不同。总之，我们发现需要根据要递送的细胞类型仔细选择纤维蛋白原和凝血酶浓度；也就是说，需要具有不同纤维蛋白原和凝血酶浓度的不同纤维蛋白构建体来单独递送成纤维细胞或角质形成细胞，或共递送两种细胞类型。此外，当它们被共同引入 3D 纤维蛋白构建体时，角质形成细胞和成纤维细胞之间似乎存在交叉对话。这种反馈可能是由于两种细胞类型在 3D 纤维蛋白构建体中产生的生长因子的作用。
