The LSK cell fraction of HSC was transduced with retrovirus-shEP4 on retronectin using ViroMag.
The paper shows the high efficiency of Magnetofection method from OZ Biosciences to improve infection and transduction capacity on hematopoietic stem cells.article reference: Blood. 2013 Mar 14;121(11):1995-2007.
Prostaglandin E2 regulates murine hematopoietic stem/progenitor cells directly via EP4 receptor and indirectly through mesenchymal progenitor cells.
Abstract
Prostaglandin E2 (PGE2) regulates hematopoietic
stem/progenitor cell (HSPC) activity. However, the receptor(s)
responsible for PGE2 signaling remains unclear. Here, we identified EP4
as a receptor activated by PGE2 to regulate HSPCs. Knockdown of Ep4 in
HSPCs reduced long-term reconstitution capacity, whereas an
EP4-selective agonist induced phosphorylation of GSK3β and β-catenin and
enhanced long-term reconstitution capacity. Next, we analyzed the
niche-mediated effect of PGE2 in HSPC regulation. Bone marrow
mesenchymal progenitor cells (MPCs) expressed EP receptors, and
stimulation of MPCs with PGE2 significantly increased their ability to
support HSPC colony formation. Among the EP receptor agonists, only an
EP4 agonist facilitated the formation of HSPC colonies after the
coculture with MPCs. PGE2 up-regulated the expression of cytokine-, cell
adhesion-, extracellular matrix-, and protease-related genes in MPCs.
We also examined the function of PGE2/EP4 signaling in the recovery of
the HSPCs after myelosuppression. The administration of PGE2 or an EP4
agonist facilitated the recovery of HSPCs from 5-fluorouracil
(5-FU)-induced myelosuppression, indicating a role for PGE2/EP4
signaling in this process. Altogether, these data suggest that EP4 is a
key receptor for PGE2-mediated direct and indirect regulation of HSPCs.
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