Adipose-derived Mesenchymal Stem Cells Therapy for Cornea Regeneration: A Multi-photon Intravital Study in a Mouse Model

Program Number: 3475 Presentation Time: 11:00 AM–11:15 AM Adipose-derived Mesenchymal Stem Cells Therapy for Cornea Regeneration: A Multi-photon Intravital Study in a Mouse Model Ladan Espandar1, Jonathan Mandell1, Tomas Blanco2. 1University of Pittsburgh, Pittsburgh, PA; 2Duke University, Durham, NC. Purpose: Adipose-derived stem cells (ADSCs) have gained increasing attention as a source of regenerative therapy because of their abundance, easy accessibility and capacity for self-renewal and differentiation; however, their fate, dynamics and differentiation of engrafted ADSCs are not well understood. Insight into in vivo stem cell biology during corneal repair could help to optimize therapeutic strategies for corneal diseases. Multiphoton intra-vital microscopy (MP-IVM) allows for noninvasive, real-time, and serial quantitative imaging in living animals. We explored the location of engrafted GFP-labeled ADSCs in mouse cornea after subconjunctival and intraperitoneal (IP) injection of cells in a total corneal epithelium debridement model by MP-IVM. Methods: Subcutaneous white adipose tissue was collected from enhanced green fluorescence protein (eGFP) transgenic mice (C57Bl/6-Tg(UBC-GFP)) and ADSCs were isolated and expanded. Eight C57Bl/6 male mice were used in the study (4 in each group). After application of ethanol (100%), the epithelium was mechanically scraped from the whole corneal surface in the right eye of each mouse. Then, in group A, GFP-mADSCs (1x106 in 50 μL) were injected subconjunctivally in both eyes (N=4). In group B, after debridement in right eye, 1x106 cells/ μL were injected IP. (N=4) MP-IVM follow up and slit-lamp examination were performed for a period of 4 weeks. Results: Subconjunctivally-injected GFP-mADSCs were found in the epithelium and anterior stroma in the injured eye by MP-IVM. No cells were found in uninjured eyes. Conversely GFP-mADSCs were found in epithelium, anterior and posterior stroma of injured cornea after IP injection. No cells were found in the contralateral uninjured eye. After 4 weeks, mice were euthanized and the results were confirmed by histology. Conclusions: ADSCs injected either subconjunctivally or IP engrafted within the cornea in response to an injury and aid to restore the damage tissue. This study opens a new promising field for the management of the ocular surface diseases by using stem cell therapy. Further studies with longer follow-ups are needed to dissect the exact functional and molecular characteristics of mADSCs in corneal wound healing process that will help to optimize possible therapeutic application of mesenchymal stem cells in corneal diseases. Commercial Relationships: Ladan Espandar, None; Jonathan Mandell, None; Tomas Blanco, None Support: NIH P30-EY08098