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A biomimetic scaffold for culturing limbal stem cells: a promising alternative for clinical transplantation
University of Ottawa Eye Institute, Ottawa, ON, Canada.
Sudhakar and Sreekant Ravi Stem Cell Laboratory, L. V. Prasad Eye Institute, Hyderabad Eye Research Foundation, Hyderabad, India.
University of Ottawa Eye Institute, Ottawa, ON, Canada and Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Canada.ORCID iD: 0000-0003-1222-6720
University of Ottawa Eye Institute, Ottawa, ON, Canada.
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2008 (English)In: JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE, ISSN 1932-6254, Vol. 2, no 5, 263-271 p.Article in journal (Refereed) Published
Abstract [en]

Limbal tissues can be cultured on various types of scaffolds to create a sheet of limbal-corneal epithelium for research as well as clinical transplantation. An optically clear, biocompatible, biomimetic scaffold would be an ideal replacement graft for transplanting limbal stem cells. in this study, we evaluated the physical and culture characteristics of the recombinant human cross-linked collagen scaffold (RHC-III scaffold) and compared it with denuded human amniotic membrane (HAM). Optical/mechanical properties and microbial susceptibility were measured for the scaffolds. With the approval of the institutional review board, 2 mm. fresh human limbal tissues were cultured on 2.5 x 2.5 cm(2) scaffolds in a medium containing autologous serum in a feeder cell-free submerged system. The cultured cell systems were characterized by morphology and immunohistochemistry for putative stem cells and differentiated cell markers. The refractive index (RI) and tensile strength of the RHC-III scaffold were comparable to human cornea, with delayed in vitro degradation compared to HAM. RHC-III scaffolds were 10-fold less susceptible to microbial growth. Cultures were initiated on day 1, expanded to form a monolayer by day 3 and covered the entire growth surface in 10 days. Stratified epithelium on the scaffolds was visualized by transmission electron microscopy. The cultured cells showed p63 and ABCG2 positivity in the basal layer and were immunoreactive for cytokeratin K3 and K12 in the suprabasal layers. RHC-III scaffold supports and retains the growth and stemness of limbal stem cells, in addition to resembling human cornea; thus, it could be a good replacement scaffold for growing cells for clinical transplantation.

Place, publisher, year, edition, pages
John Wiley and Sons , 2008. Vol. 2, no 5, 263-271 p.
Keyword [en]
tissue engineering; replacement grafts; recombinant collagen cross-linked scaffolds; biomimetic; limbal stem cells; explant culture; transplantation; biocompatible
National Category
Engineering and Technology
URN: urn:nbn:se:liu:diva-70658DOI: 10.1002/term.91ISI: 000257807100003OAI: diva2:441081
Available from: 2011-09-14 Created: 2011-09-14 Last updated: 2013-12-17

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Griffith, May
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