Fetal Fibroblasts and Keratinocytes with Immunosuppressive Properties for Allogeneic Cell-Based Wound Therapy
Authors
Thomas Zuliani, Soraya Saiagh, Anne-Chantal Knol, Julie Esbelin, Brigitte Dreno
Institution
INSERM, Nantes
Country
France
Year
2013
Journal
PLOS
Abstract
Fetal skin heals rapidly without scar formation early in gestation, conferring to fetal skin cells a high and unique potential for
tissue regeneration and scar management. In this study, we investigated the possibility of using fetal fibroblasts and
keratinocytes to stimulate wound repair and regeneration for further allogeneic cell-based therapy development. From a
single fetal skin sample, two clinical batches of keratinocytes and fibroblasts were manufactured and characterized.
Tolerogenic properties of the fetal cells were investigated by allogeneic PBMC proliferation tests. In addition, the potential
advantage of fibroblasts/keratinocytes co-application for wound healing stimulation has been examined in co-culture
experiments with in vitro scratch assays and a multiplex cytokines array system. Based on keratin 14 and prolyl-4-
hydroxylase expression analyses, purity of both clinical batches was found to be above 98% and neither melanocytes nor
Langerhans cells could be detected. Both cell types demonstrated strong immunosuppressive properties as shown by the
dramatic decrease in allogeneic PBMC proliferation when co-cultured with fibroblasts and/or keratinocytes. We further
showed that the indoleamine 2,3 dioxygenase (IDO) activity is required for the immunoregulatory activity of fetal skin cells.
Co-cultures experiments have also revealed that fibroblasts-keratinocytes interactions strongly enhanced fetal cells
secretion of HGF, GM-CSF, IL-8 and to a lesser extent VEGF-A. Accordingly, in the in vitro scratch assays the fetal fibroblasts
and keratinocytes co-culture accelerated the scratch closure compared to fibroblast or keratinocyte mono-cultures. In
conclusion, our data suggest that the combination of fetal keratinocytes and fibroblasts could be of particular interest for
the development of a new allogeneic skin substitute with immunomodulatory activity, acting as a reservoir for wound
healing growth factors.