McIntyre A P, Bjornson B H
Exp Hematol. 1986 Oct;14(9):833-9.
Long-term bone marrow cultures are dependent on the formation in vitro of an adherent cell layer that supports hematopoiesis. We have grown bone-marrow-adherent cells, termed stromal colony-forming units, or CFU-ST, as isolated adherent colonies, and examined some of their growth requirements. Bone marrow mononuclear cells separated from aspirates by density centrifugation and cultured in medium supplemented with fetal calf serum or human plasma gave rise to adherent colonies (CFU-ST). An average of 23.4 +/- 2.1 (mean +/- SEM, n = 19) CFU-ST were produced by 10(5) bone marrow mononuclear cells. CFU-ST could not be cultured from similarly prepared peripheral blood mononuclear cells. The colonies were composed of spindle cells, flat cells, and fat-containing cells, with all three types often present in the same colony, suggesting derivation from a common progenitor. Cells were negative for nonspecific esterase and factor VIII antigen. Hydrocortisone added to the cultures at concentrations of 10(-7) M induced the formation of adipose cells in the center of one-third to one-half of the colonies but did not affect CFU-ST number. Human platelet-poor plasma and platelet-rich plasma were substituted for fetal calf serum in the medium. When all determinations for four experiments were averaged, platelet-rich plasma gave 17.8 +/- 1.2 (mean +/- SEM, n = 16) colonies, whereas platelet-poor plasma gave only 0.2 +/- 0.1 colonies (n = 15). When purified platelet-derived growth factor (PDGF) was added to platelet-poor plasma, growth of CFU-ST was enhanced, and a dose-response relationship was found between size of colonies and concentration of added PDGF. Granulocyte-macrophage colony stimulating factor added to cultures had no effect on the growth of CFU-ST.
长期骨髓培养依赖于体外形成支持造血的贴壁细胞层。我们已将称为基质集落形成单位（CFU-ST）的骨髓贴壁细胞培养成分离的贴壁集落，并研究了它们的一些生长需求。通过密度离心从骨髓抽吸物中分离出的骨髓单个核细胞，在补充有胎牛血清或人血浆的培养基中培养，可形成贴壁集落（CFU-ST）。10⁵个骨髓单个核细胞平均产生23.4±2.1（均值±标准误，n = 19）个CFU-ST。从同样制备的外周血单个核细胞中无法培养出CFU-ST。集落由纺锤形细胞、扁平细胞和含脂肪细胞组成，这三种类型的细胞常同时存在于同一个集落中，提示它们来源于共同的祖细胞。细胞对非特异性酯酶和因子VIII抗原呈阴性。以10⁻⁷M的浓度添加到培养物中的氢化可的松，可诱导三分之一至二分之一的集落中心形成脂肪细胞，但不影响CFU-ST的数量。用人少血小板血浆和富血小板血浆替代培养基中的胎牛血清。当对四个实验的所有测定结果进行平均时，富血小板血浆产生17.8±1.2（均值±标准误，n = 16）个集落，而少血小板血浆仅产生0.2±0.1个集落（n = 15）。当将纯化的血小板衍生生长因子（PDGF）添加到少血小板血浆中时，CFU-ST的生长得到增强，并且发现集落大小与添加的PDGF浓度之间存在剂量反应关系。添加到培养物中的粒细胞-巨噬细胞集落刺激因子对CFU-ST的生长没有影响。
