Zuckerman K S, Wicha M S
Blood. 1983 Mar;61(3):540-7.
We have studied the deposition of extracellular matrix proteins in the adherent stroma of long-term murine bone marrow cultures. Stable hematopoiesis was maintained for greater than 12 wk. At selected intervals, culture dishes were sacrificed by removing all nonadherent cells and air drying the dishes. The adherent stromal layer was analyzed for the presence of intracellular and extracellular collagen, fibronectin, and laminin using double immunofluorescent staining with specific antisera against these matrix components. In cultures examined during the first 2 wk, large numbers of stromal cells contained collagen, fibronectin, and laminin. Over the next 2 wk, an extensive extracellular network of fibronectin, laminin, and collagen was deposited on the dishes, which persisted throughout the life of the cultures. In contrast to a previous report, we detected substantial numbers of endothelial cells by means of immunofluorescent staining of stromal cells with antisera to type IV collagen, laminin, and factor VIII antigen. Although deposition of these extracellular matrix proteins coincides with onset of active hematopoietic cell production, the relative roles of the stromal cells and the extracellular matrix in supporting hematopoiesis in murine bone marrow cell cultures remain to be determined.
我们研究了长期小鼠骨髓培养物贴壁基质中细胞外基质蛋白的沉积情况。稳定的造血功能维持了超过12周。在选定的时间间隔，通过去除所有非贴壁细胞并将培养皿风干来牺牲培养皿。使用针对这些基质成分的特异性抗血清进行双重免疫荧光染色，分析贴壁基质层中细胞内和细胞外胶原蛋白、纤连蛋白和层粘连蛋白的存在情况。在最初2周内检查的培养物中，大量基质细胞含有胶原蛋白、纤连蛋白和层粘连蛋白。在接下来的2周内，大量由纤连蛋白、层粘连蛋白和胶原蛋白组成的细胞外网络沉积在培养皿上，并在培养物的整个生命周期中持续存在。与之前的一份报告相反，我们通过用抗IV型胶原蛋白、层粘连蛋白和因子VIII抗原的抗血清对基质细胞进行免疫荧光染色，检测到了大量内皮细胞。尽管这些细胞外基质蛋白的沉积与活跃造血细胞产生的开始同时发生，但基质细胞和细胞外基质在支持小鼠骨髓细胞培养中的造血作用仍有待确定。
