Enhanced viability of corneal epithelial cells for efficient transport/storage using a structurally modified calcium alginate hydrogel

Stem Cells & Nanomaterials Laboratory, Reading School of Pharmacy, Hopkins Building, University of Reading, Berkshire, RG6 6UB, UK.
Regenerative Medicine (Impact Factor: 2.79). 05/2012; 7(3):295-307. DOI: 10.2217/rme.12.7
Source: PubMed


Therapeutic limbal epithelial stem cells could be managed more efficiently if clinically validated batches were transported for 'on-demand' use.
In this study, corneal epithelial cell viability in calcium alginate hydrogels was examined under cell culture, ambient and chilled conditions for up to 7 days.
Cell viability improved as gel internal pore size increased, and was further enhanced with modification of the gel from a mass to a thin disc. Ambient storage conditions were optimal for supporting cell viability in gel discs. Cell viability in gel discs was significantly enhanced with increases in pore size mediated by hydroxyethyl cellulose.
Our novel methodology of controlling alginate gel shape and pore size together provides a more practical and economical alternative to established corneal tissue/cell storage methods.

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Available from: Che John Connon, Sep 17, 2014
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    • "We have previously demonstrated that calcium alginate gels are suitable for storage/transport of immobilized embryonic stem cells, mesenchymal stem cells, and LESC [20], [22], proving that these 3-D matrices are capable of retaining the progenitor phenotype of encapsulated cells. "
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    ABSTRACT: Limbal epithelial stem cells may ameliorate limbal stem cell deficiency through secretion of therapeutic proteins, delivered to the cornea in a controlled manner using hydrogels. In the present study the secretome of alginate-encapsulated limbal epithelial stem cells is investigated. Conditioned medium was generated from limbal epithelial stem cells encapsulated in 1.2% (w/v) calcium alginate gels. Conditioned medium proteins separated by 1-D gel electrophoresis were visualized by silver staining. Proteins of interest including secreted protein acidic and rich in cysteine, profilin-1, and galectin-1 were identified by immunoblotting. The effect of conditioned medium (from alginate-encapsulated limbal epithelial stem cells) on corneal epithelial cell proliferation was quantified and shown to significantly inhibit (P≤0.05) their growth. As secreted protein acidic and rich in cysteine was previously reported to attenuate proliferation of epithelial cells, this protein may be responsible, at least in part, for inhibition of corneal epithelial cell proliferation. We conclude that limbal epithelial stem cells encapsulated in alginate gels may regulate corneal epithelialisation through secretion of inhibitory proteins.
    PLoS ONE 07/2013; 8(7):e70860. DOI:10.1371/journal.pone.0070860 · 3.23 Impact Factor
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    • "Because the structure of alginate gels can be modified to direct cell phenotype, particularly stem cell differentiation [57,58], and enhance cell survival, this biomaterial presents an ideal culture system for LEC, which may be developed for application to reversing the pathology of LSCD. Moreover, the well-reported use of alginate gels for cell preservation [59,60] will allow this hydrogel to be developed into a transport and/or storage medical device (Fig. 3) applicable to corneal cell-based therapies. "
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    ABSTRACT: Corneal blindness caused by limbal stem cell deficiency (LSCD) is a prevailing disorder worldwide. Clinical outcomes for LSCD therapy using amniotic membrane (AM) are unpredictable. Hydrogels can eliminate limitations of standard therapy for LSCD, because they present all the advantages of AM (i.e. biocompatibility, inertness and a biodegradable structure) but unlike AM, they are structurally uniform and can be easily manipulated to alter mechanical and physical properties. Hydrogels can be delivered with minimum trauma to the ocular surface and do not require extensive serological screening before clinical application. The hydrogel structure is also amenable to modifications which direct stem cell fate. In this focussed review we highlight hydrogels as biomaterial substrates which may replace and/or complement AM in the treatment of LSCD.
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    ABSTRACT: The administration of probiotic bacteria as nutraceuticals is an area that has rapidly expanded in recent years, with a global market worth $32.6 billion predicted by 2014. Many of the health promoting claims attributed to these bacteria are dependent on the cells being both viable and sufficiently numerous in the intestinal tract. The oral administration of most bacteria results in a large loss of viability associated with passage through the stomach, which is attributed to the high acid and bile salt concentrations present. This loss of viability effectively lowers the efficacy of the administered supplement. The formulation of these probiotics into microcapsules is an emerging method to reduce cell death during GI passage, as well as an opportunity to control release of these cells across the intestinal tract. The majority of this technology is based on the immobilization of bacteria into a polymer matrix, which retains its structure in the stomach before degrading and dissolving in the intestine, unlike the diffusion based unloading of most controlled release devices for small molecules. This review shall provide an overview of progress in this field as well as draw attention to areas where studies have fallen short. This will be followed by a discussion of emerging trends in the field, highlighting key areas in which further research is necessary.
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