Multipotent hematopoietic progenitor cells immortalized by Lhx2 self-renew by a cell nonautonomous mechanism
Multipotent hematopoietic progenitor cells immortalized by Lhx2 self-renew by a cell nonautonomous mechanism
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DOI：
10.1016/s0301-472x(01)00666-x
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发表时间：
2001-08-01
影响因子：
2.6
通讯作者：
Carlsson, L
中科院分区：
文献类型：
--
作者：
do O, PP;Wandzioch, E;Carlsson, L
Objective.造血干细胞（HSCs）的直接分子和细胞研究受到造血组织中低水平HSCs的阻碍。为了解决这些问题，我们通过在衍生自体外分化的胚胎干细胞的造血祖细胞中表达LIM同源框基因Lhx 2（先前为LH 2）来产生永生化多能造血前体细胞（HPC）系。为了进一步验证HPC系作为正常HSC模型的相关性，我们详细分析了HPC系体外生长的要求。Lhx 2通过一种新颖的细胞非自主机制使HPC系永生化。HPC系的自我更新依赖于功能性Lhx 2表达。大多数早期作用的造血相关生长因子对HPC系表现出协同作用，而晚期作用因子本身不诱导分化。转化生长因子-β（1）是HPC系增殖的有效抑制剂。HPC细胞株接种于基质细胞株后，可高效形成鹅卵石区域，且鹅卵石区域形成细胞可在基质细胞株中维持数月。我们的数据显示，在许多方面，HPC系在细胞水平上类似于正常的造血祖细胞/干细胞，与大多数先前描述的多能造血细胞系相反。细胞非自主机制的永生化的HPC线表明，Lhx 2调节，直接或间接，可溶性介质参与的HPC线的自我更新。(C)2001年国际实验血液学学会。出版社：Elsevier Science Inc.
Objective. Direct molecular and cellular studies of hematopoietic stem cells (HSCs) are hampered by the low levels of HSCs in hematopoietic tissues. To address these issues, we generated immortalized multipotent hematopoietic precursor cell (HPC) lines by expressing the LIM-homeobox gene Lhx2 (previously LH2) in hematopoietic progenitors derived from embryonic stem cells differentiated in vitro.Materials and Methods. To validate further the relevance of the HPC lines as a model for normal HSCs, we analyzed in detail the growth requirements of HPC lines in vitro.Results. Lhx2 immortalized the HPC lines by a putatively novel and cell nonautonomous mechanism. Self-renewal of the HPC lines is dependent on functional Lhx2 expression. Most early-acting hemato poiesis-related growth factors show synergistic effects on the HPC lines, whereas late-acting factors do not induce differentiation by themselves. Transforming growth factor-beta (1) is a potent inhibitor of proliferation of the HPC lines. HPC lines form cobblestone areas with high efficiency when seeded onto stromal cell lines, and the cobblestone area-forming cell can be maintained in these cultures for several months.Conclusions. Our data show that, in many respects, HPC lines are similar to normal hematopoietic progenitor/stem cells on the cellular level, in contrast to most previously described multipotent hematopoietic cell lines. The cell nonautonomous mechanism for immortalization of the HPC lines suggests that Lhx2 regulates, directly or indirectly, soluble mediators involved in self-renewal of the HPC lines. (C) 2001 International Society for Experimental Hematology. Published by Elsevier Science Inc.