Characterization of corneal stromal stem cells with the potential for epithelial transdifferentiation

Hashmani, Khurram; Branch, Matthew J.; Sidney, Laura E.; Dhillon, Permesh Singh; Verma, Megha; McIntosh, Owen D.; Hopkinson, Andrew; Dua, Harminder S.

doi:10.1186/scrt226

Cited by 76 publications

(66 citation statements)

References 81 publications

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“…2F). Earlier studies, based on FACS analysis, reported CD29 as a marker of mesenchymal-like cells of the anterior limbal stroma [22,23,26]. However, we observed that the epithelial cells of individual holoclones specifically expressed CD29, and the colony periphery was lined by nestin-positive cells (Fig.…”

Section: Spatial Distribution Of Limbal Niche Cells In Vitromentioning

confidence: 46%

Spatial Distribution of Niche and Stem Cells in Ex Vivo Human Limbal Cultures

Mariappan

Kacham

Purushotham

et al. 2014

Stem Cells Translational Medicine

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Stem cells at the limbus mediate corneal epithelial regeneration and regulate normal tissue homeostasis. Ex vivo cultured limbal epithelial transplantations are being widely practiced in the treatment of limbal stem cell deficiency. In this report, we examined whether the limbal niche cells that nurture and regulate epithelial stem cells coexist in ex vivo limbal cultures. We also compared the inherent differences between explant and suspension culture systems in terms of spatial distribution of niche cells and their effect on epithelial stem cell proliferation, migration, and differentiation in vitro. We report that the stem cell content of both culture systems was similar, explaining the comparable clinical outcomes reported using these two methods. We also showed that the niche cells get expanded in culture and the nestin-positive cells migrate at the leading edges to direct epithelial cell migration in suspension cultures, whereas they are limited to the intact niche in explant cultures. We provide evidence that C/EBPd-positive, p15-positive, and quiescent, label-retaining, early activated stem cells migrate at the leading edges to regulate epithelial cell proliferation in explant cultures, and this position effect is lost in early suspension cultures. However, in confluent suspension cultures, the stem cells and niche cells interact with each another, migrate in spiraling patterns, and self-organize to form three-dimensional niche-like compartments resembling the limbal crypts and thereby reestablish the position effect. These 3D-sphere clusters are enriched with nestin-, vimentin-, S100-, and p27-positive niche cells and p15-, p21-, p63a-, C/EBPd-, ABCG2-, and Pax6-positive quiescent epithelial stem cells. STEM CELLS

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Section: Spatial Distribution Of Limbal Niche Cells In Vitromentioning

confidence: 46%

Spatial Distribution of Niche and Stem Cells in Ex Vivo Human Limbal Cultures

Mariappan

Kacham

Purushotham

et al. 2014

Stem Cells Translational Medicine

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“…Human corneal stromal stem cells (CSSC) were isolated from corneal rims, using a modification of a previously described method [23]. Excess sclera was removed and the epithelium and endothelium detached by gentle scraping.…”

Section: Isolation and Culture Of Primary Human Corneal Stromal Stem mentioning

confidence: 99%

“…In vitro, keratocytes extracted from the limbus, have been shown to display characteristics of multipotent mesenchymal stromal cells (MSC) [20,22], that after several passages in a certain medium, conform to a criteria stipulated by the International Society for Cellular Therapy (ISCT) [18,23]. The extracted corneal stroma-derived stem cells (CSSC) express MSC-associated cell surface markers such as CD29, CD73, CD90 and CD105, and possess the ability to differentiate down the osteogenic, chondrogenic and adipogenic lineages in vitro [18,24].…”

Section: Introductionmentioning

confidence: 99%

“…It has also been suggested that serum-free growth media developed for other cell types might also be suitable, such as the use of keratinocyte serum free medium (K-SFM) [34]. Changing the basal medium from DMEM to medium 199 (M199) or DMEM/F12, which contain a greater proportion of other components such as amino acids and nucleotides leads to the generation of a population of MSC or MSC-like cells [23,35]. The use of M199 with the addition of 20% FBS to culture keratocytes, generates MSC that adhere to ISCT criteria [18].…”

Section: Introductionmentioning

confidence: 99%

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Effect of culture medium on propagation and phenotype of corneal stroma–derived stem cells

et al. 2015

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(2015) Effect of culture medium on propagation and phenotype of corneal stroma-derived stem cells. Cytotherapy, 17 (12). pp. 1706 -1722 . ISSN 1477 -2566 Access from the University of Nottingham repository: http://eprints.nottingham.ac.uk/31287/1/Submission%20Manuscript.pdf Copyright and reuse:The Nottingham ePrints service makes this work by researchers of the University of Nottingham available open access under the following conditions. This article is made available under the University of Nottingham End User licence and may be reused according to the conditions of the licence. For more details see: http://eprints.nottingham.ac.uk/end_user_agreement.pdf A note on versions:The version presented here may differ from the published version or from the version of record. If you wish to cite this item you are advised to consult the publisher's version. Please see the repository url above for details on accessing the published version and note that access may require a subscription. However, the optimal culture conditions are disputed and few direct media comparisons have been performed. In this report, we evaluated several media types to identify the optimal for inducing an in vitro stem cell phenotype. Discussion: Culture medium can significantly influence CSSC phenotype. SCM produced a cell phenotype closest to that of a pluripotent stem cell, and we considerate to be the most appropriate for development as a clinical grade medium for the production of CSSC phenotypes suitable for cell therapy. Methods

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“…Examples include human mesenchymal stem cells [13], human embryonic stem cells [14], human corneal stroma stem cells [15,16], adipose-derived stem cells [17,18], and dental pulp stem cells [2].…”

Section: Introductionmentioning

confidence: 99%