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Human pluripotent stem cell-derived limbal epithelial stem cells on bioengineered matrices for corneal reconstruction
BioMediTech, University of Tampere, Tampere, Finland.
Faculty of Medicine, University of Szeged, Szeged, Hungary.
LinkoCare Life Sciences AB.
Faculty of Medicine, University of Szeged, Szeged, Hungary.
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2016 (Engelska)Ingår i: Experimental Eye Research, ISSN 0014-4835, Vol. 146, s. 26-34Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Corneal epithelium is renewed by limbal epithelial stem cells (LESCs), a type of tissue-specific stem cells located in the limbal palisades of Vogt at the corneo-scleral junction. Acute trauma or inflammatory disorders of the ocular surface can destroy these stem cells, leading to limbal stem cell deficiency (LSCD) – a painful and vision-threatening condition. Treating these disorders is often challenging and complex, especially in bilateral cases with extensive damage. Human pluripotent stem cells (hPSCs) provide new opportunities for corneal reconstruction using cell-based therapy. Here, we investigated the use of hPSC-derived LESC-like cells on bioengineered collagen matrices in serum-free conditions, aiming for clinical applications to reconstruct the corneal epithelium and partially replace the damaged stroma. Differentiation of hPSCs towards LESC-like cells was directed using small-molecule induction followed by maturation in corneal epithelium culture medium. After four to five weeks of culture, differentiated cells were seeded onto bioengineered matrices fabricated as transparent membranes of uniform thickness, using medical-grade porcine collagen type I and a hybrid cross-linking technology. The bioengineered matrices were fully transparent, with high water content and swelling capacity, and parallel lamellar microstructure. Cell proliferation of hPSC-LESCs was significantly higher on bioengineered matrices than on collagen-coated control wells after two weeks of culture, and LESC markers p63 and cytokeratin 15, along with proliferation marker Ki67 were expressed even after 30 days in culture. Overall, hPSC-LESCs retained their capacity to self-renew and proliferate, but were also able to terminally differentiate upon stimulation, as suggested by protein expression of cytokeratins 3 and 12. We propose the use of bioengineered collagen matrices as carriers for the clinically-relevant hPSC-derived LESC-like cells, as a novel tissue engineering approach for corneal reconstruction.

Ort, förlag, år, upplaga, sidor
Academic Press, 2016. Vol. 146, s. 26-34
Nyckelord [en]
Corneal epithelium;Limbal epithelial stem cells;Human pluripotent stem cells;Serum-free culture conditions;Tissue engineering;Regenerative medicine
Nationell ämneskategori
Cell- och molekylärbiologi
Identifikatorer
URN: urn:nbn:se:liu:diva-125224DOI: 10.1016/j.exer.2015.11.021ISI: 000376332400005OAI: oai:DiVA.org:liu-125224DiVA, id: diva2:903654
Anmärkning

Funding agencies:  LinkoCare Life Sciences AB; Academy of Finland [218050, 133879]; Finnish Funding Agency for Technology and Innovation (TEKES); Doctoral Programme in Biomedicine and Biotechnology; Eye and Tissue Bank Foundation; Emil Aaltonen Foundation; Foundation Suppor

Tillgänglig från: 2016-02-16 Skapad: 2016-02-16 Senast uppdaterad: 2018-01-10

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Ratnayake, AnjulaRafat, Mehrdad

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Cell- och molekylärbiologi

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