Optimization of optical and mechanical properties of real architecture for 3-dimensional tissue equivalents: Towards treatment of limbal epithelial stem cell deficiency

Massie, Isobel; Kureshi, Alvena; Schrader, Stefan; Shortt, Alex J.; Daniels, Julie T.

doi:10.1016/j.actbio.2015.06.007

Cited by 30 publications

(29 citation statements)

References 41 publications

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“…RAFT TEs could then be used as a delivery vehicle for transplantation of HLE. In addition, clinically appropriate collagen can be used and its density can be tailored to provide optimal transparency and mechanical strength of the tissue equivalents 30 .…”

Section: Discussionmentioning

confidence: 99%

Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents

Kureshi

Dziasko

Funderburgh

et al. 2015

Sci Rep

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Human limbal epithelial cells (HLE) and corneal stromal stem cells (CSSC) reside in close proximity in vivo in the corneal limbal stem cell niche. However, HLE are typically cultured in vitro without supporting niche cells. Here, we re-create the cell-cell juxtaposition of the native environment in vitro, to provide a tool for investigation of epithelial-stromal cell interactions and to optimize HLE culture conditions for potential therapeutic application. RAFT (Real Architecture For 3D Tissue) tissue equivalents (TEs) were used as a 3-dimensional substrate for co-culturing HLE and CSSC. Our results demonstrate that a monolayer of HLE that maintained expression of p63α, ABCB5, CK8 and CK15 (HLE markers), formed on the surface of RAFT TEs within 13 days of culture. CSSC remained in close proximity to HLE and maintained expression of mesenchymal stem cell markers. This simple technique has a short preparation time of only 15 days with the onset of HLE layering and differentiation observed. Furthermore, co-cultivation of HLE with another niche cell type (CSSC) directly on RAFT TEs, eliminates the requirement for animal-derived feeder cells. RAFT TEs may be useful for future therapeutic delivery of multiple cell types to restore the limbal niche following ocular surface injury or disease.

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Section: Discussionmentioning

confidence: 99%

Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents

Kureshi

Dziasko

Funderburgh

et al. 2015

Sci Rep

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“…Some LESC supports are regularly used in the clinic, such as human amniotic membrane (HAM) or fibrin gel . A number of other supports are in preclinical development and may even offer better standardization and reproducibility than the clinically used ones . Most often, cells are cultured on denuded HAM , which is essentially a basement membrane with a composition similar to the limbal epithelial basement membrane .…”

Section: Stem Cells For Epithelial Wound Healingmentioning

confidence: 99%

Concise Review: Stem Cells for Corneal Wound Healing

et al. 2017

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Corneal wound healing is a complex process that occurs in response to various injuries and commonly used refractive surgery. It is a significant clinical problem, which may lead to serious complications due to either incomplete (epithelial) or excessive (stromal) healing. Epithelial stem cells clearly play a role in this process, whereas the contribution of stromal and endothelial progenitors is less well studied. The available evidence on stem cell participation in corneal wound healing is reviewed, together with the data on the use of corneal and non‐corneal stem cells to facilitate this process in diseased or postsurgical conditions. Important aspects of corneal stem cell generation from alternative cell sources, including pluripotent stem cells, for possible transplantation upon corneal injuries or in disease conditions are also presented. Stem Cells 2017;35:2105–2114

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“…“ Real Architecture For 3D Tissue” (RAFT™) combines the advantages of ECM collagen composition with three dimensional expansion. RAFT™ recently has been introduced in the research field of human ophthalmology as an ex vivo tissue culture system for culturing limbal fibroblast (Massie et al 2015a), limbal epithelial cells (Massie et al 2015b) and for improved interaction of corneal stem cells with limbal epithelial cells (Kureshi et al 2015). Here we report, for the first time how RAFT™ tissue culture platform preserves pancreatic islet morphology, aids islet viability and supports insulin and glucagon production.…”

Section: Introductionmentioning

confidence: 99%

Real architecture For 3D Tissue (RAFT™) culture system improves viability and maintains insulin and glucagon production of mouse pancreatic islet cells

Szebeni¹,

Táncos

Feher³

et al. 2017

Cytotechnology

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There is an unmet medical need for the improvement of pancreatic islet maintenance in culture. Due to restricted donor availability it is essential to ameliorate islet viability and graft engraftment. The aim of this study was to compare the standard tissue culture techniques with the advanced Real Architecture For 3D Tissue (RAFT™) culture system in terms of viability and hormone production. Here, we first report that islets embedded in RAFT™ collagen type I advanced tissue culture system maintain their tissue integrity better than in monolayer and suspension cultures. The Calcein violet assay and Annexin V/propidium-iodide staining show higher cell viability in the RAFT™ culture system. Quantitative real-time PCR data showed that RAFT™ increases insulin expression after 18 days in culture compared to traditional methods. Enhanced insulin and glucagon production was further verified by immunofluorescent staining in a time-course manner. These results indicate that RAFT™ tissue culture platform can be a promising tool to maintain pancreatic islet spheroid integrity and culture islets for downstream high throughput pharmacological studies ex vivo.Electronic supplementary materialThe online version of this article (doi:10.1007/s10616-017-0067-6) contains supplementary material, which is available to authorized users.

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Optimization of optical and mechanical properties of real architecture for 3-dimensional tissue equivalents: Towards treatment of limbal epithelial stem cell deficiency

Abstract: Graphical abstract

Cited by 30 publications

References 41 publications

Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents

Human corneal stromal stem cells support limbal epithelial cells cultured on RAFT tissue equivalents

Concise Review: Stem Cells for Corneal Wound Healing

Real architecture For 3D Tissue (RAFT™) culture system improves viability and maintains insulin and glucagon production of mouse pancreatic islet cells

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