Horikoshi A, Murphy M J
Exp Cell Biol. 1984;52(6):371-82. doi: 10.1159/000163284.
In an attempt to more closely simulate the native hematopoietic environment, human bone marrow bony matrix was cultivated in long-term human bone marrow cultures. Sternal bone marrow curettings (i.e., 'bony matrix') were cultured with and without autologous bone marrow single cell suspensions. Fresh media were provided at weekly intervals and the 'harvested' cells were assayed for CFU-gm (i.e., the granulocyte-macrophage precursor). Bone marrow bony matrix alone was not competent to maintain CFU-gm production, although early in the culture nucleated cells and CFU-gm were recruited from the bony matrix. A dose of 850 rad of X-radiation to the bone marrow matrix damaged the hematopoietic growth-promoting effects of the adherent cell layer, which was, however, rapidly reconstituted by an inoculation of intact autologous bone marrow cells. Additional investigations revealed that, whereas the addition of hydrocortisone to this culture system did not alter the maintenance of CFU-gm, recharging the cultures with fresh autologous marrow maintained CFU-gm for 10 weeks. These data indicate that human bone marrow stroma is more sensitive to X-irradiation in vitro than in vivo, and the bone marrow hematopoietic microenvironment provided by its bony matrix seemed to be less effective in vitro than in vivo with respect to the maintenance of hematopoietic precursor cells.
为了更紧密地模拟天然造血环境，在长期人骨髓培养中培养人骨髓骨基质。将胸骨骨髓刮除物（即“骨基质”）与自体骨髓单细胞悬液一起或不与自体骨髓单细胞悬液一起培养。每周更换新鲜培养基，并对“收获”的细胞进行CFU-gm（即粒细胞-巨噬细胞前体）检测。单独的骨髓骨基质不能维持CFU-gm的产生，尽管在培养早期有核细胞和CFU-gm从骨基质中募集。对骨髓基质进行850拉德的X射线照射会破坏贴壁细胞层的造血生长促进作用，然而，通过接种完整的自体骨髓细胞可迅速重建该作用。进一步的研究表明，在该培养系统中添加氢化可的松不会改变CFU-gm的维持情况，用新鲜自体骨髓补充培养物可使CFU-gm维持10周。这些数据表明，人骨髓基质在体外比在体内对X射线照射更敏感，并且就维持造血前体细胞而言，其骨基质提供的骨髓造血微环境在体外似乎不如在体内有效。
