Abstract
To reduce the increasing need for corneal transplantation, attempts are currently aiming to restore corneal clarity, one potent source of cells are multipotent adult progenitor cells (MAPC®). These cells release a powerful cocktail of paracrine factors that can guide wound healing and tissue regeneration. However, their role in corneal regeneration has been overlooked. Thus, we sought to explore the potential of combining the cytoprotective storage feature of alginate, with MAPC to generate a storable cell-laden gel for corneal wound healing. 72 hours following hypothermic storage, alginate encapsulation was shown to maintain MAPC viability at either 4 or 15°C. Encapsulated MAPC (2 x106 cells/mL) stored at 15°C presented the optimum temperature that allowed for cell recovery. These cells had the ability to reattach to tissue culture plastic whilst exhibiting normal phenotype and this was maintained in serum-free and xenobiotic-free medium. Furthermore, corneal stromal cells presented a significant decrease in scratch-wounds in the presence of alginate encapsulated MAPC compared to a no-cell control (p = 0.018). This study shows that immobilization of MAPC within an alginate hydrogel does not hinder their ability to affect a secondary cell population via soluble factors and that these effects are successfully retained following hypothermic storage.
Highlights
Corneal damage and opacity have been estimated to cause blindness in 8 million people (c.10% of total blindness) worldwide each year [1]
Within this study, we investigated the potential of alginate-encapsulation of multipotent adult progenitor cells (MAPC) to maintain cell viability, morphology, and ability to affect the closure of an in vitro corneal stromal scratch-wound via paracrine factors following 72 hours of hypothermic storage at 4 and 15 ̊C
Starting with exploring the effect of XFM media on MAPC cultures phenotype, MAPC thawed from liquid nitrogen seeded in a 2000 cells/cm2 density and grown and maintained in XFM medium for 72 hours at 37 ̊C were analyzed for morphology, proliferation, and viability
Summary
Corneal damage and opacity have been estimated to cause blindness in 8 million people (c.10% of total blindness) worldwide each year [1]. Corneal keratocytes usually remain quiescent and are only activated when penetrating damage to the tissue occurs [3]. Multipotent adult progenitor cell in corneal stromal wound healing does not alter our adherence to PLOS ONE policies on sharing data and protocols. Very few cell-based therapy products have been approved by the European Medicines Agency and the Food and Drug Administration (FDA) [4,5,6,7,8]
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