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Susumu Sakimoto, Valentina Marchetti, Edith Aguilar, Yoshihiko Usui, Daniel Feitelberg, Carli M Wittgrove, Jennifer K Trombley, Martin Friedlander; Endothelial Colony Forming Cells derived in Xeno-free Media Facilitate Retinal Vascular Repair in Oxygen Induced Retinopathy. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1827.
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© ARVO (1962-2015); The Authors (2016-present)
The human cord blood derived endothelial colony forming cell (ECFC) has emerged as a promising target for vascular regenerative therapies. However, in vitro expansion of ECFCs depends on the presence of fetal bovine serum (FBS) in the culture media, a component that is not appropriate for clinical application in humans. Moreover, the precise role of ECFCs in ocular ischemic diseases has not yet been clarified. This study evaluated vascular repair in a mouse model of oxygen-induced retinopathy (OIR) using ECFCs derived in xeno free media (XFM) or in serum-containing media (SCM).
ECFCs were derived in XFM or SCM. In vitro morphology, colony forming capability, cell surface antigen expression, and acetylated LDL uptake were evaluated. To evaluate in the OIR model, at p17 retinas were imaged and analyzed for obliteration area and neovascularization.
Colonies derived in either media displayed a cobblestone appearance with variations in size, consistent with their heterogeneous proliferative rates previously reported. The number of colonies detected 10 days after plating were 13.8[CP1] in XFM and 20.0 in SCM (n=3). ECFCs derived and amplified in either media expressed endothelial markers (CD31 and CD144), did not express hematopoietic markers (CD45), and took up equivalent amounts acetylated LDL. In the OIR model, injection of ECFCs derived and amplified in either XFM or SCM decreased the area of vascular obliteration and neovascular tufts to a similar level compared to vehicle injection (p=0.019 and p=0.018, respectively in one-way analysis of variance).
We demonstrate that ECFCs in XFM displayed robust clonal proliferative ability in vitro. They facilitate vascular repair in OIR to the same extent of which conventionally derived ECFCs do. These data suggest that ECFCs exhibit a novel therapeutic approach for the treatment of ischemic retinopathies and that the use of xeno-free media will facilitate the isolation and proliferation of ECFCs for use in the clinics.
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