Murine Adherent CD^4+ Cell Population Expanded by Single Cell Cloning

Introduction: while human endothelial progenitor cells (epcs) have been asubject of somehow extensive investigation, epcs from adult mousehematopoietic system were poorly studied. present investigation is focusedon fvb mouse endothelial progenitor cells in terms of their isolation,purification, and expansion.material and methods: mononuclear cells collected from murine peripheralblood were cultured in fibronectin coated plate for two weeks, at which point,the adherent cell population were lifted and analyzed in terms of somesurface markers. using facs vantage equipped with one-cell depositionunit, single cd34 positive cells were plated per well already containingmedium optimized for single cell growth. several clones were then emerged,expanded, and examined in terms of some surface markers. furthermore,the cells were investigated regarding ability to uptake dil-ac-ldl and formcapillary network on matrigel surfaces.results: adherent population of mononuclear cells from mouse peripheralblood was appeared morphologically heterogeneous. about 5% of theadherent cells were cd34 positive. having optimized their culture condition,several cd34 positive clones were expanded. the cells comprising theclones were dil-ac-ldl+ and formed capillary-like tube when being seededon matrigel surfaces.conclusion: the primary culture of the mononuclear cells from murineperipheral blood contains a very limited number of cells positive forendothelial lineage markers. these cells (adherent cd34 positive) could beexpanded by single cell cloning technique