Lim B, Izaguirre C A, Aye M T, Huebsch L, Drouin J, Richardson C, Minden M D, Messner H A
J Cell Physiol. 1986 Apr;127(1):45-54. doi: 10.1002/jcp.1041270107.
The maintenance of hemopoietic precursors in long-term liquid bone marrow cultures (LTBMC) is associated with the presence of an adherent stromal layer composed of heterogeneous cell populations. We have used a culture assay to promote the growth of one of its cellular components and characterize its properties. Freshly obtained bone marrow cells and cells derived from the adherent layer of LTBMC were grown in methylcellulose-clotted plasma in the presence of phytohemagglutinin-stimulated leukocyte-conditioned medium (PHA-LCM), hydrocortisone (HC), and citrated normal human plasma. Both sources contained cells (CFU-RF) that gave rise to colonies of cells with a reticulofibroblastoid appearance. In the presence of HC, most colonies contained lipid-laden cells. Colonies could be further propagated as adherent layers when transferred into liquid cultures. These cells produced laminin, fibronectin, and collagen types I, III, IV, and V. They were negative for Von Willebrand factor VIII. The ability to synthesize laminin and collagen type IV distinguished these cells from a population of previously described bone marrow fibroblasts (CFU-F). The relationship of CFU-RF to hemopoietic precursors was investigated using patients with chronic myeloid leukemia and bone marrow transplant recipients. Cells within CFU-RF-derived colonies were uniformly negative for the Philadelphia chromosome, thus making it unlikely that they belonged to the malignant hemopoietic clone. CFU-RF-derived colonies in bone marrow transplant recipients were found to be exclusively of host origin. Both observations support the view that CFU-RF is not part of the repertoire of hemopoietic stem cells.
长期液体骨髓培养（LTBMC）中造血前体细胞的维持与由异质细胞群体组成的贴壁基质层的存在有关。我们使用了一种培养测定法来促进其细胞成分之一的生长并表征其特性。将新鲜获得的骨髓细胞和源自LTBMC贴壁层的细胞在植物血凝素刺激的白细胞条件培养基（PHA-LCM）、氢化可的松（HC）和枸橼酸化正常人血浆存在的情况下，在甲基纤维素凝结的血浆中培养。这两种来源都含有能产生具有网状纤维母细胞样外观的细胞集落的细胞（CFU-RF）。在HC存在的情况下，大多数集落含有充满脂质的细胞。当转移到液体培养中时，集落可以作为贴壁层进一步传代。这些细胞产生层粘连蛋白、纤连蛋白以及I、III、IV和V型胶原蛋白。它们对血管性血友病因子VIII呈阴性。合成层粘连蛋白和IV型胶原蛋白的能力将这些细胞与先前描述的骨髓成纤维细胞群体（CFU-F）区分开来。使用慢性髓性白血病患者和骨髓移植受者研究了CFU-RF与造血前体细胞的关系。CFU-RF衍生集落内的细胞对费城染色体均呈阴性，因此它们不太可能属于恶性造血克隆。在骨髓移植受者中发现CFU-RF衍生的集落完全来自宿主。这两个观察结果都支持CFU-RF不是造血干细胞库一部分的观点。
