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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
ZUSAMMENFASSUNG

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.

MATERIALIEN
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Produktbeschreibung

Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, for molecular biology
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, ACS reagent, ≥99.5%
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Essigsäure, glacial, ACS reagent, ≥99.7%
Sigma-Aldrich
Essigsäure, glacial, ReagentPlus®, ≥99%
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Trypanblau -Lösung, 0.4%, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Ethyl alcohol, Pure, 200 proof, meets USP testing specifications
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
Formaldehyd -Lösung, for molecular biology, 36.5-38% in H2O
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Essigsäure, glacial, ≥99.99% trace metals basis
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L-Glutamin, meets USP testing specifications, suitable for cell culture, 99.0-101.0%, from non-animal source
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Propidiumjodid, ≥94.0% (HPLC)
SAFC
Formaldehyd -Lösung, contains 10-15% methanol as stabilizer, 37 wt. % in H2O
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Essigsäure -Lösung, suitable for HPLC
Sigma-Aldrich
Essigsäure, glacial, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, 99.8-100.5%
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Essigsäure, glacial, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8%
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Ethanol, BioUltra, for molecular biology, ≥99.8%, (absolute alcohol, without additive, A15 o1)
Sigma-Aldrich
L-Glutamin, ReagentPlus®, ≥99% (HPLC)
Sigma-Aldrich
Ethanol, ACS reagent, prima fine spirit, without additive, F15 o1