Moreau I, Andreoni C, Caux C, Saeland S, Rigal D
Immunology Laboratory, C.R.T.S., Lyon, France.
Eur J Haematol. 1992 Jul;49(1):29-35. doi: 10.1111/j.1600-0609.1992.tb00910.x.
Differences in the plastic adhesive properties of bone marrow (BM) cells were used to initiate modified stromal layers (MSL) from long-term cultures by removing non-adherent cells shortly (4 to 18 hours) after initial seeding. Following this early modification, adherent cells generated a confluent layer after 21 days of culture. Cellular characteristics of volume and spontaneous fluorescence determined by flow cytometry showed that the MSL included 82% fibroblastic stromal cells, 8% macrophages and 10% myelomonocytic cells. Furthermore, clonogenic assays revealed that the MSL were devoid of hematopoietic progenitor cells. MSL were found to sustain long-term myelopoiesis for at least 7 weeks from exogenously added hematopoietic progenitors isolated from bone marrow (CD34+ cells), thereby demonstrating their functionality. The present experimental model appears of interest for the study of interactions between defined populations of hematopoietic cells and cells of the adherent layer. Of importance, our present modifications of human long-term bone marrow culture are technically simple and do not involve manipulation of the stromal cells.
利用骨髓（BM）细胞的可塑性黏附特性，在初次接种后不久（4至18小时）去除非黏附细胞，从而从长期培养物中起始改良基质层（MSL）。经过这种早期改良后，贴壁细胞在培养21天后形成汇合层。通过流式细胞术测定的细胞体积和自发荧光特征表明，MSL包含82%的成纤维细胞样基质细胞、8%的巨噬细胞和10%的髓单核细胞。此外，集落形成试验显示MSL缺乏造血祖细胞。发现MSL能从外源性添加的从骨髓分离的造血祖细胞（CD34+细胞）维持至少7周的长期髓系造血，从而证明了它们的功能。本实验模型似乎对研究特定造血细胞群体与贴壁层细胞之间的相互作用很有意义。重要的是，我们目前对人类长期骨髓培养的改良技术简单，且不涉及对基质细胞的操作。
