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Qi Yan, Qiru Wang, Xanthi Couroucli; HPP-EPC Characterization And The Potential Role In Retinopathy Of Prematurity. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3183.
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Retinopathy of prematurity (ROP) is a disease that affects immature vasculature in the eyes of preterm infants. It can become aggressive with retinal neovascularization and progress to retinal detachment and blindness. As smaller babies are surviving, the incidence of ROP has increased. With the tremendous therapeutic potential of endothelial progenitor cells (EPC) in the treatment of ischemic diseases, the goal of this study is to identify and characterize a subset of functional HPP-EPC from bone marrow. We will further test their ability to promote vascular regrowth and repair aberrant retinal vessels in ROP model.
To enhance endothelial differentiation, an in vitro model with BMEC-CM was developed to induce bone marrow-derived EPC. Endothelial cell colonies (EC-cols) were cultured with BMEC-CM for 3 months. EC markers and functions, the leukocytes marker CD45 and monocytic functions were studied. The single progenitor cell growth curve was quantified. The high-proliferative potential (HPP)-EPC were isolated and characterized.
BMEC-CM treatment of bone marrow mononuclear cells showed the best induction, with EPC-colony cell expression of CD31, von Willebrand factor, UEA-1 positive, and incorporation of Ac-LDL. EPC-colony cells expressed differential proliferative capacity; the HPP-EPC were isolated, they exhibited a high proliferative capacity up to 20 population doublings resulting in 7x10^6 cells per EPC after 14 weeks incubation, and formed the capillary-like network in vitro. Moreover, HPP-EPC expressed CD45, exhibited endocytic activities, and secrete various growth factors into the culture medium.
HPP-EPC were isolated and cultured with the induction of BMEC-CM. These cells showed high proliferative capacity and preserved some hematopoietic lineage characteristics. We will further test their ability to facilitate functional vessel growth in ROP model.
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