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Howard Ray, Janice Park, Jennifer Mansour, Scott Seaman, Bijan Dey, Shayn Peirce, Paul Andrew Yates; A Lineage Tracing System to Fate Map Perivascular-Derived Adipose Stem Cells in Retinal Vasculopathy Mouse Models. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1831.
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© ARVO (1962-2015); The Authors (2016-present)
The application of adipose-derived stem cells (ASCs) as a cellular therapy for vascular retinopathy is highly promising, however the exact cell of origin for ASCs remains in dispute and to-date no genetic lineage analysis has been performed. The purpose of this work was to characterize myosin heavy chain 11 (Myh11) expressing perivascular cells in vivo and perivascular-derived adipose stem cells (ASCs) harvested from a tamoxifen inducible lineage tracing mouse model ("Myh11 eYFP+/+"). We demonstrate Myh11 eYFP+ perivascular cells constitute a subpopulation of ASCs that can be intravitreally injected and fluorescently tracked in retinal vasculopathy mouse models.
Epididymal and inguinal adipose tissue from adult tamoxifen induced Myh11 eYFP+/+ mice were harvested and examined under direct fluorescence microscopy as well as immunofluorescently stained to determine Myh11 expression via eYFP signal. Confocal imaging and ImageJ analysis was used to determine eYFP signal overlap with the pericyte marker, PDGFRβ. Epididymal and inguinal adipose tissue was digested with collagenase to investigative the eYFP signal within the stromal vascular fraction (SVF) and ASC population in vitro.
Expression of eYFP was found throughout the perivascular region of epididymal and inguinal adipose tissue, which was verified with lectin co-labeling (Figure 1). After image analysis, eYFP expression was found to partially co-localize with PDGFRβ in both adipose tissues. From flow cytometry analysis on the SVF, we found that eYFP positive cells represent less than 5% of the entire SVF population. After plating and passaging the SVF on tissue culture plastic, eYFP cells continue to express their marker and constitute less than 10% of cells found in vitro.
The Myh11 eYFP+/+ mouse model can provide a valuable means to fate map in vivo injected eYFP+ perivascular-derived ASCs in retinal vasculopathy mouse models. We expect to intravitreally inject these cells within retinal vasculopathy mouse models to determine their cell fate as well as efficacy of this sub-population of ASCs on the retinal vasculature and neural retina.
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