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  • Bovine posterior limbus: an evaluation of an alternative source for corneal endothelial and trabecular meshwork stem/progenitor cells.

Bovine posterior limbus: an evaluation of an alternative source for corneal endothelial and trabecular meshwork stem/progenitor cells.

Stem cells and development (2014-10-18)
Wing Yan Yu, Ian Grierson, Carl Sheridan, Amy Cheuk-Yin Lo, David Sai-Hung Wong
RESUMO

A growing body of evidence has revealed that stem-like cells in the posterior limbus of the eye between the corneal endothelium (CE) and trabecular meshwork (TM) may be able to rejuvenate these tissues in disease. However, these cells have not been clearly defined and we have named them PET cells (progenitor cells of the endothelium and trabeculum). A good and inexpensive animal model for PET cells is lacking, so we investigated bovine eyes as an effective large tissue source. We showed the presence of stem/progenitor cells in the bovine CE, transition zone, and TM in situ. Floating spheres cultured from the CE and TM showed similar stem cell marker expression patterns. Both the CE and TM spheres were bipotent and highly proliferative, but with limited secondary sphere-forming capability. They were highly prone to differentiate back into the cell type of their tissue of origin. It is speculated that the PET cells become more tissue-specific as they migrate away from their niche. Here, we showed that PET cells are present in the posterior limbus of bovine eyes and that they can be successfully cultured and expanded. PET cells represent an attractive target for developing new treatments to regenerate both the CE and TM, thereby reducing the requirement for donor tissue for corneal transplant and invasive treatments for glaucomatous patients.

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Álcool etílico, puro, 200 proof, for molecular biology
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Álcool etílico, puro, 200 proof, ACS reagent, ≥99.5%
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Ácido acético, glacial, ACS reagent, ≥99.7%
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Ácido acético, glacial, ReagentPlus®, ≥99%
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Álcool etílico, puro, 200 proof, HPLC/spectrophotometric grade
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Azul de triptano, 0.4%, liquid, sterile-filtered, suitable for cell culture
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Álcool etílico, puro, 200 proof, meets USP testing specifications
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DAPI, for nucleic acid staining
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Álcool etílico, puro, 190 proof, for molecular biology
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Formaldeído, for molecular biology, 36.5-38% in H2O
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Ácido acético, glacial, ≥99.99% trace metals basis
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L-Glutamina, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
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Iodeto de propídio, ≥94.0% (HPLC)
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Álcool etílico, puro, 200 proof, anhydrous, ≥99.5%
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Acetic acid solution, suitable for HPLC
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Etanol, BioUltra, for molecular biology, ≥99.8%, (absolute alcohol, without additive, A15 o1)
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L-Glutamina, ReagentPlus®, ≥99% (HPLC)