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Involvement of hematopoietic stem cells in liver fibrosis : clarification of origin of hepatic stellate cells using single cell transplantation method

造血干细胞参与肝纤维化：利用单细胞移植方法阐明肝星状细胞的起源

基本信息

批准号：
18591055
负责人：
MASUYA Masahiro
金额：
$ 2.53万
依托单位：
Mie University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

We have known that non-blood cells, which are derived from hematopoietic stem cells, exist in the CCl_4-injured liver ; however, we have never evaluated the characteristics of the cells. Single Lin-c-kit^+Sca-l^+CD34^- cells from EGFP mice (EGFP^+ KSL-CD34^- cells) were deposited into round-bottomed 96-well plates and cultured in the presence of IL-11 and SCF for 7 days. The contents of the wells containing 20 or fewer viable clusters of cells derived from single EGFP^+KSL-CD34^- cells were injected into the tail veins of irradiated C57BL/6J mice. We induced liver injury by CCl_4 treatment in transplanted mice, which showed high levels of long-term, multilineage hematopoietic engraftment at 2 months after BM transplantation. After 12 weeks of CCl_4 treatment, we detected EGFP^+ hepatic stellate cells, which contained intracytoplasmic lipid droplets(Oil-red-O positive) and expressed vimentin, glial fibrillary acidic protein, and ADAMTS 13. Moreover, EGFP^+ cells isolated from the liver … More synthesized collagen I in culture. The hematopoietic stem cell-derived hepatic stellate cells seen in male-to-male transplants revealed only one Y chromosome. We presented our research results in 2006 ASH annual meeting (at Orlando) and published them in Blood Journal(2008 ; 111 : 2427-2435).Although we proved that hematopoietic stem cells can transdifferentiate into hepatic stellate cells in liver injury, pathways and mechanisms that regulate the transdifferentiation are remained unknown. We have found that myeloid lineage cells, especially monocytes and macrophages, can transdifferentiate into glomerular mesangial cells in our previous study. Then, we tried to examine the relationship between myeloid lineage cells and hepatic stellate cells. Monocytes and macrophages were prepared by the culture of EGFP^+ KSL cells in the presence of SCF and M-CSF, and injected into liver-injured mice. We detected that the number of EGFP^+ cells in these mice were similar to those in bone marrow total nucleated cells-transplanted mice. Now, we are evaluating whether the EGFP^+ cells are Kupffer cells or hepatic stellate cells. Less

我们已经知道在CCl_4损伤的肝脏中存在来源于造血干细胞的非血细胞，但我们从未对这些细胞的特性进行过评估。取绿色荧光蛋白小鼠单个LIN-c-kit^+Sca-L^+CD34^-细胞(EGFP^+KSL-CD34^-细胞)接种于96孔板中，加入IL-11和SCF培养7天。将单个EGFP^+KSL-CD34^-细胞中含有20个或更少活性细胞簇的孔中的内容物注入照射后的C57BL/6J小鼠尾静脉。我们用CCl_4诱导了移植小鼠的肝损伤，在骨髓移植后2个月表现出高水平的长期多系造血植入。CCl_4处理12周后，我们检测到EGFP^+的肝星状细胞，其中含有胞浆内的脂滴(油红-O阳性)，并表达波形蛋白、胶质纤维酸性蛋白和ADAMTS13。此外，从肝脏…分离的EGFP^+细胞在培养中合成更多的I型胶原。在男性对男性移植中看到的造血干细胞来源的肝星状细胞只显示了一条Y染色体。我们在2006年奥兰多ASH年会上发表了我们的研究成果，并发表在血液杂志(2008；111：2427-2435)上。尽管我们证明了造血干细胞在肝损伤中可以转分化为肝星状细胞，但调节转分化的途径和机制仍不清楚。我们在之前的研究中发现，髓系细胞，特别是单核细胞和巨噬细胞，可以转分化为肾小球系膜细胞。然后，我们试图研究髓系细胞和肝星状细胞之间的关系。将EGFP~+KSL细胞在SCF和M-CSF存在下培养，制备单核细胞和巨噬细胞，注射入肝损伤小鼠体内。我们检测到这些小鼠的EGFP^+细胞数量与骨髓全有核细胞移植小鼠相似。现在，我们正在评估EGFP^+细胞是Kupffer细胞还是肝星状细胞。较少