The 5-lipoxygenase pathway regulates vasculogenesis in differentiating mouse embryonic stem cells

作者：Finkensieper Andreas^*; Kieser Sophia; Bekhite Mohamed M; Richter Madeleine; Mueller Joerg P; Graebner Rolf; Figulla Hans Reiner; Sauer Heinrich; Wartenberg Maria

来源：Cardiovascular Research, 2010, 86(1): 37-44.

DOI：10.1093/cvr/cvp385

• Embryonic stem cells
• Embryoid body
• Leukotrienes
• 5-Lipoxygenase
• 5-Lipoxygenase activating protein

It is well established that leukotrienes (LTs), products of the 5-lipoxygenase (5-LO) pathway, participate in inflammatory tissue reactions and immune responses. In the present study, the impact of the 5-LO pathway on vasculogenesis of mouse embryonic stem (ES) cells was investigated.
Immunohistochemistry studies demonstrated that 5-LO( ) cells first appeared at day 6 of embryoid body (EB) formation from ES cells. 5-LO( )/CD68( ) as well as 5-LO( )/CD45( ) cells were prominent at day 10 of EB differentiation. Real-time PCR and western blot analysis revealed all constituents of the 5-LO pathway. High performance liquid chromatography analyses indicated the synthesis of LTB(4) and LTD(4) in conformity with induction of the 5-LO pathway. Furthermore, Flk-1( )/CD105( ) cells displayed calcium- (Ca(2 )) transients in response to LTB(4), whereas CD11b( ) cells responded to LTD(4). Treatment of EBs with LTB(4) and LTD(4) resulted in phosphorylation of the mitogen-activated protein kinase ERK1/2. Pharmacological inhibition of the 5-LO pathway and stable shRNA targeting of 5-LO-activating protein decreased capillary cell areas positive for PECAM-1.
Our data demonstrate that the 5-LO pathway emerges early during ES cell differentiation into cells of the myeloid lineage and that LTs play an until now unrecognized role in vascular development of ES cells.

• 出版日期2010-4-1