Wang T Y, Brennan J K, Wu J H
University of Rochester, Department of Chemical Engineering, NY 14627-0166.
Exp Hematol. 1995 Jan;23(1):26-32.
The highly packed cell density and the three-dimensional structure in the hematopoietic compartment of bone marrow facilitate cell-to-cell and cell-to-matrix interactions known to be important for hematopoietic activities. To provide a similar environment in vitro, we developed a long-term bone marrow culture (LTBMC) system, continuously perfused with Dexter's medium, employing packed, highly porous bovine collagen microspheres as the matrix support for marrow cell growth. Using murine bone marrow as a model, we found that the culture system differed from the conventional flask culture in the following ways: 1) as revealed by the electron microscopy, the bone marrow cells in the culture system grew in a three-dimensional configuration, similar to that in vivo, 2) the cell output from the culture system at 37 degrees C was virtually the same as that at 33 degrees C, and 3) in the absence of exogenous growth factors, except those in the serum, the culture system produced lymphoid cells and all stages of committed cells (i.e., erythrocytes, granulocytes, macrophages, and megakaryocytes), thus indicating multilineal differentiation of the hematopoietic stem cells. Furthermore, cell clusters resembling erythroblastic islands were observed in the absence of exogenous erythropoietin (Epo). The culture system appears to provide a different microenvironment than that of the flask culture and may be used as an alternative model for hematopoiesis.
骨髓造血区中高度密集的细胞密度和三维结构促进了细胞间以及细胞与基质间的相互作用，而这些相互作用已知对造血活动很重要。为了在体外提供类似的环境，我们开发了一种长期骨髓培养（LTBMC）系统，该系统持续灌注德克斯特培养基，使用紧密堆积的、高度多孔的牛胶原微球作为骨髓细胞生长的基质支持物。以小鼠骨髓为模型，我们发现该培养系统与传统的培养瓶培养在以下方面存在差异：1）如电子显微镜所示，培养系统中的骨髓细胞以三维结构生长，类似于体内情况；2）该培养系统在37℃时的细胞输出量与33℃时几乎相同；3）在没有外源性生长因子（血清中的生长因子除外）的情况下，该培养系统产生淋巴细胞以及各阶段的定向分化细胞（即红细胞、粒细胞、巨噬细胞和巨核细胞），这表明造血干细胞具有多系分化能力。此外，在没有外源性促红细胞生成素（Epo）的情况下，观察到了类似成红细胞岛的细胞簇。该培养系统似乎提供了一种与培养瓶培养不同的微环境，可作为造血的替代模型。
