Purchase this article with an account.
Juan Liang, Vickie Hoi Ying Wong, Lynsey-Dawn Allen, Stephen J Elliman, Paul Loftus, Lisa O'Flynn, Alan W Stitt; CD362+ stromal stem cells(SSCs) stabilize vascular networks through differentiation into pericyte-like cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6076.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Cell-based therapy provides a promising strategy to address pericyte loss and microvascular insufficiency in early diabetic retinopathy. In our previous study, a sub-population of bone marrow-derived stromal stem cells (SSCs) sorted on CD362 promoted vascular repair in oxygen-induced retinppathy (OIR). The present work focus on the related mechanism to study angiogenesis in 3D culture system.
SSCs were sorted from human bone marrow mononuclear cells on the basis of the presence or absence of surface marker CD362 (syndecan-2). This created two distinct populations of SSCs described as CD362+ and CD362- cells. In vitro tubulogenesis was conducted under hypoxia(1% O2) using two inter-related approaches: 1) A Matrigel assay using endothelial colony forming cells (ECFCs ) (7x104) mixed with CD362+SSCs (1X104) were suspended in Matrigel which was spotted onto 24 well plates; 2) A double-layer Matrigel assay in which ECFCs were suspended in Matrigel and spotted onto 24 well plates followed by topping of another Matrigel layer containing CD362+SSCs one day later. Tube-like structures were imaged using confocal microscopy or conducted for immunostaining assay.
The pericyte markers, NG2, PDGFR β, α-SMA were detected in both CD362+ SSCs and CD362-SSCs, in which significant increase of α-SMA expression cells were found in CD362+ SSCs compared to CD362- SSCs (45.7% ±0.04 in CD 362+SSCs and 24.5% ±0.06 in CD362-SSCs, p=0.0306,n=4 ). In the in vitro angiogenesis assay, regression of ECFC tubules occurred at 1 week while the presence of CD362+SSCs stabilized the network with the cells taking on a perivascular location. CD362+SSCs /ECFC co-culture maintained stabilized vascular networks until two weeks by showing significantly enhanced tube area(p=0.005,n=4) and branch points (p<0.0001,n=8) compared to ECFCs alone. In the double-layer Matrigel assay, CD362+SSCs migrated into “pre-formed” ECFC networks and took up a perivascualr position. Preicyte markers were expressed in those CD362+SSCs resident alongside tubes.
CD362+ SSCs appeared to act as pericyte progenitors by showing the capacity to associate with vascular tubes and stabilize the network under hypoxia conditions. These cells have promise for cell-replacement therapy for retinal ischemic diseases such as diabetic retinopathy.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
This PDF is available to Subscribers Only