Murine hepatocyte cell lines promote expansion and differentiation of NK cells from stem cell precursors
Murine hepatocyte cell lines promote expansion and differentiation of NK cells from stem cell precursors
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DOI：
10.1002/hep.20234
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发表时间：
2004-06-01
期刊：
影响因子：
13.5
通讯作者：
Tripodi, M
中科院分区：
文献类型：
--
作者：
Bordoni, V;Alonzi, T;Tripodi, M
虽然胎儿肝脏是主要的造血器官，但正常成人肝脏为各种免疫细胞提供了合适的微环境，并且在几种病理条件下，可能成为髓外造血的部位。肝细胞对造血细胞分化的直接影响知之甚少。我们以前曾报道过，Met鼠肝细胞（MMH）未转化的肝细胞系保留了肝细胞在体内的几个形态和功能特征，并能够支持细胞-细胞接触系统中造血前体的生存，自我更新和分化。在这里，我们报告的MMH系释放的可溶性因子对骨髓源性细胞的影响。在两种不同的无细胞接触系统中培养造血干细胞：MMH饲养层上的transwell插入物和MMH条件培养基（MMH-CM）。两种培养系统都能够促进骨髓源性细胞的大量扩增及其向表达NK1.1和U 5A 2 -13标志物的自然杀伤（NK）细胞的分化。纯化的造血干细胞（Sca-1 + Lin-），无论是作为一个整体群体或作为单细胞接种，也能够分化成NK细胞，当在MMH-CM中培养时，因此，MMH系分泌的可溶性因子促进NK前体细胞的扩增和分化。MMH-CM衍生的NK细胞具有功能活性;需要通过白细胞介素（IL）-12和IL-18的刺激来诱导干扰素-γ（IFN-γ）表达并增强其细胞毒性活性。总之，我们的研究结果可能意味着肝细胞在NK细胞发育中的直接作用，我们所使用的系统可能为研究NK细胞分化的分子机制提供工具。
While fetal liver is a major hematopoietic organ, normal adult liver provides a suitable microenvironment for a variety of immune cells and, in several pathological conditions, may become a site of extramedullary hematopoiesis. The direct influence of hepatocytes on hematopoietic cell differentiation is poorly understood. We have previously reported that the Met murine hepatocyte (MMH) untransformed hepatocytic lines retain several morphological and functional features of hepatocytes in vivo and are able to support the survival, self-renewal, and differentiation of hematopoietic precursors in a cell-cell contact system. Here we report the effects of soluble factors released by MMH lines on bone marrow-derived cells. Lymphohematopoietic cells were cultured in two different cell contact-free systems: transwell inserts on MMH feeder layers, and MMH conditioned medium (MMH-CM). Both culture systems were able to promote a substantial expansion of bone marrow-derived cells and their differentiation to natural killer (NK) cells that express the NK1.1 and U5A2-13 markers. Purified hematopoietic stem cells (Sca-1 + Lin-), either plated as a bulk population or as single cells, were also able to differentiate into NK cells, when cultured in MMH-CM; thus, soluble factors secreted by MMH lines promote the expansion and differentiation of NK precursor cells. MMH-CM-derived NK cells are functionally active; stimulation by interleukin (IL)-12 together with IL-18 was required to induce interferon-gamma (IFNgamma) expression and to enhance their cytotoxic activity. In conclusion our findings may imply a direct role of hepatocytes in NK cell development, and the system we have used may provide a tool for studying the molecular mechanisms of NK cell differentiation.