Koguma M, Matsuda K, Okuyama R, Yanai N, Obinata M
Department of Cell Biology, Institute of Development, Aging, and Cancer, Tohoku University, Japan.
Exp Hematol. 1998 Apr;26(4):280-7.
To understand the regulatory mechanisms involved in the development of hematopoietic stem cells, we cultured lineage-negative, c-Kit+ Sca-1+ stem cells sorted from bone marrow cells by a fluorescence-activated cell sorter (FACS) on layers of bone marrow stromal cell lines established from SV40 T-antigen gene transgenic mice. We previously reported that the TBR59 stromal cell line induced two sequential cobblestone formations: the first formation committed to the myeloid and the second to the lymphoid lineage. After examination of many other bone marrow stromal cell lines, we found that TBR31-1 stromal cells supported only lymphoid development of the sorted stem cells. The sorted stem cells proliferated by forming cobblestones and the cells were released from the cobblestones. Most released cell populations were B220-positive lymphoid cells; cell production continued for 2 months. Addition of G-CSF or M-CSF produced only a slight effect on myeloid development. FACS analysis of the released cells showed that the B-lymphoid-committed progenitors developed into mature B-cells by expressing surface immunoglobulin M. These results indicate that TBR31-1 bone marrow stromal cells selectively support B-lymphoid development, whereas TBR59 cells support both myeloid and lymphoid development of stem cells.
为了解造血干细胞发育过程中的调控机制，我们将通过荧光激活细胞分选仪（FACS）从骨髓细胞中分选出来的谱系阴性、c-Kit+ Sca-1+干细胞，培养在由SV40 T抗原基因转基因小鼠建立的骨髓基质细胞系层上。我们之前报道过，TBR59基质细胞系诱导了两个连续的鹅卵石样集落形成：第一个集落形成致力于髓系，第二个致力于淋巴系。在检测了许多其他骨髓基质细胞系后，我们发现TBR31-1基质细胞仅支持分选的干细胞的淋巴系发育。分选的干细胞通过形成鹅卵石样集落进行增殖，并且细胞从鹅卵石样集落中释放出来。大多数释放的细胞群体是B220阳性淋巴细胞；细胞产生持续了2个月。添加粒细胞集落刺激因子（G-CSF）或巨噬细胞集落刺激因子（M-CSF）对髓系发育仅产生轻微影响。对释放细胞的FACS分析表明，B淋巴细胞定向祖细胞通过表达表面免疫球蛋白M发育成成熟B细胞。这些结果表明，TBR31-1骨髓基质细胞选择性地支持B淋巴细胞发育，而TBR59细胞支持干细胞的髓系和淋巴系发育。
