Mechanotransductive Differentiation of Hair Follicle Stem Cells Derived from Aged Eyelid Skin into Corneal Endothelial-Like Cells

Abstract

Corneal endothelial insufficiency is one of the leading causes of blindness. The main contemporary treatment for corneal blindness is endothelial keratoplasty, which, however, is unsatisfactory as a medical therapy due to the lack of donor corneas and graft rejection. Therefore, autologous stem cell-based corneal endothelial tissue substitutes may be a promising alternative to conventional grafts in the future. To address the age of most patients suffering from corneal endothelial deficiencies, we investigated the presence and potential of hair-derived stem cells from older tissue donors. Our studies revealed the presence of pluripotency- and neural crest-associated markers in tissue sections from blepharoplasty patients aged 50 to 80 years. In vitro outgrowths from eyelid hair follicles on collagen-coated tissue culture plates revealed a weak decrease in stem-cell potency. In contrast, cells within the spheres that spontaneously formed from the adherent cell layer retained full stem-cell potency and could be differentiated into cells of the ecto- meso and endodermal lineages. Although these highly potent hair follicle derived stem cells (HFSC) were only very slightly expandable, they were able to recognize the biomimicry of the Descemet’s-like topography and differentiate into corneal endothelial-like cells. In conclusion, HFSCs derived from epidermal skin of eyelid biopsies are a promising cell source to provide autologous corneal endothelial replacement for any age group of patients. Graphical Abstract

Safety and efficacy of human ESC-derived corneal endothelial cells for corneal endothelial dysfunction

“…The results demonstrated that the differentiated iCECs on day 24 were mature iCECs. Our data presented here support prior reports that the membrane potential of cells can be measured by the combination of oscilloscope and Transwells to evaluate cell maturity [ 31 ]. Moreover, RNA-seq data identified potential molecular markers of mature iCECs, which may serve as surrogates for predicting their functional properties.…”

“…Given that TJP1 (ZO-1) , CDH2 (N-cadherin) , ATP1A1 (Na + K + ATPase) and AQP1 play critical roles in the function of CECs, we hypothesize that a prolonged maturation culture might be needed for functional acquisition of iCECs. To demonstrate the functional capacity of iCECs, we performed transendothelial electrical potential difference recording of day 19 and day 24 cells using a Transwell system as described previously [ 31 ]. This transendothelial potential is dependent on continuous tight junctions and the activity of Na + K + ATPase, which is of fundamental importance for the osmotic activity of iCECs.…”

Adult stem cells in the eye: Identification, characterisation, and therapeutic application in ocular regeneration – A review

“…135 The hair follicular stem cells are multipotent, capable of giving rise to not only the follicle but also the epidermis and the sebaceous gland. 136 Studies showed that hair follicular stem cells could be transdifferentiated into cells that resemble the corneal epithelium and endothelium, 137,138 demonstrating great promise for ocular regeneration.…”

Recent advances in cell-based regenerative therapies for corneal disease

“…Meanwhile, additional candidates for adjunctive therapy are being investigated, such as fibroblast grow factor 1 derivatives which may promote endothelial proliferation and migration [61,62], and other preclinical studies use cells that will not require corneal donation, such as human SC from embryonic [63], induced pluripotent [64–67], umbilical cord-derived [68], dental pulp [69] and eyelid hair follicle-derived [70] sources. None of these approaches are currently in human trials.…”