Establishment of induced pluripotent stem cells from aged mice using bone marrow-derived myeloid cells
Establishment of induced pluripotent stem cells from aged mice using bone marrow-derived myeloid cells
复制标题
DOI：
10.1093/jmcb/mjq044
复制
发表时间：
2011-04-01
影响因子：
5.5
通讯作者：
Isobe, Ken-ichi
中科院分区：
文献类型：
--
作者：
Cheng, Zhao;Ito, Sachiko;Isobe, Ken-ichi
如果要将诱导多能干细胞（iPS）用于老年患者的损伤组织治疗或器官修复，就必须从老年患者的组织中培养iPS细胞。为了确定在老年患者中使用这项技术的可行性，我们询问是否确实有可能从老年小鼠的骨髓（BM）中建立iPS细胞。将携带绿色荧光蛋白（GFP）基因的老年C57BL/6小鼠骨髓细胞与粒细胞巨噬集落刺激因子（GM-CSF）一起培养4天。将四种因子（Oct3/4、Sox2、Klf4和c-Myc）引入脑源性髓细胞（BM-M）。在GM-CSF中培养的衰老脑脊髓瘤生成iPS细胞的效率较低。然而，我们成功地从携带GFP的衰老C57BL/6小鼠中获得了BM-M-iPS细胞。我们的BM-M-iPS细胞表达SSEA-1和Pou5f1，碱性磷酸酶染色呈阳性。诱导多能干细胞注射到同基因C57BL/6小鼠体内后，确实形成了三胚层畸胎瘤，并在体外分化为三胚层。通过与OP9共培养，BM-M-iPS细胞可以向髓系分化。分化后的BM-M-iPS细胞在GM-CSF存在下增殖良好，Nanog和Pou5f1的表达缺失，至少部分原因是其启动子甲基化。相反，Tnf和Il1b基因表达上调，其启动子低甲基化。
If induced pluripotent stem (iPS) cells are to be used to treat damaged tissues or repair organs in elderly patients, it will be necessary to establish iPS cells from their tissues. To determine the feasibility of using this technology with elderly patients, we asked if it was indeed possible to establish iPS cells from the bone marrow (BM) of aged mice. BM cells from aged C57BL/6 mice carrying the green fluorescence protein (GFP) gene were cultured with granulocyte macrophage-colony stimulating factor (GM-CSF) for 4 days. Four factors (Oct3/4, Sox2, Klf4 and c-Myc) were introduced into the BM-derived myeloid (BM-M) cells. The efficiency of generating iPS cells from aged BM cultured in GM-CSF was low. However, we succeeded in obtaining BM-M-iPS cells from aged C57BL/6 mice, which carried GFP. Our BM-M-iPS cells expressed SSEA-1 and Pou5f1 and were positive for alkaline phosphatase staining. The iPS cells did make teratoma with three germ layers following injection into syngeneic C57BL/6 mice, and can be differentiated to three germ layers in vitro. By co-culturing with OP9, the BM-M-iPS cells can be differentiated to the myeloid lineage. The differentiated BM-M-iPS cells proliferated well in the presence of GM-CSF, and lost expression of Nanog and Pou5f1, at least in part, due to methylation of their promoters. On the contrary, Tnf and Il1b gene expression was upregulated and their promoters were hypomethylated.