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LIM Homeobox Domain 2 Is Required for Corneal Epithelial Homeostasis.

Sartaj R, Chee RI, Yang J, Wan P, Liu A, Guaiquil V, Fuchs E, Rosenblatt MI - Stem Cells (2016)

Bottom Line: Immunodetection on corneal sections were used to visualize conjunctivalization, a sign of limbal barrier failure.Cell based assays showed that Lhx2cKO derived corneal epithelial cells have a significantly lower capacity to form colonies over time and delayed wound-healing recovery when compared to wildtype cells.We conclude that Lhx2 is required for maintenance of the corneal epithelial cell compartment and the limbal barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus

Lhx2cKO derived corneal epithelial cells showed delayed wound healing.Corneal epithelial cells were isolated from WT and Lhx2cKO mice and cultured as indicated in Material and Methods. Cells were seeded in culture plates and when confluent a scratch was made through the cell monolayer and the wound closure followed via time‐lapse microscopy (A). Scratches in WT cells closed in less than 24 hrs, while closure of scratches made in Lhx2cKO cultures were significantly delayed, healing by 48hrs (B). Data are means SE ± (n = 3). *P ≤ 0.05. Abbreviation: WT, wild type.
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stem2257-fig-0004: Lhx2cKO derived corneal epithelial cells showed delayed wound healing.Corneal epithelial cells were isolated from WT and Lhx2cKO mice and cultured as indicated in Material and Methods. Cells were seeded in culture plates and when confluent a scratch was made through the cell monolayer and the wound closure followed via time‐lapse microscopy (A). Scratches in WT cells closed in less than 24 hrs, while closure of scratches made in Lhx2cKO cultures were significantly delayed, healing by 48hrs (B). Data are means SE ± (n = 3). *P ≤ 0.05. Abbreviation: WT, wild type.

Mentions: Tissue injury is known to activate stem cells to proliferate and form new tissue in various adult organs 17, 18, 19, 20. In culture, we mimicked the corneal wound healing procedure by scratching confluent cultures of the WT and Lhx2cKO murine CECs and monitored the closure of the wound. Images were taken at 0, 8, 16, 24, and 48 hours at the same locations in the cultures using phase‐contrast microscopy. The cell‐free area of the scratch in WT and Lhx2cKO cells was measured at time 0 and selected for direct comparisons only if they had approximately the same wound area between replicas to monitor healing over time. We found a significant decrease in Lhx2cKO wound closure when compared with WT (Fig. 4A). Quantification of the cell‐free areas showed that Lhx2cKO cells delay wound healing as early as 8 hours (2.2 ± 1.16) when compared with the WT (7.5 ± 1.46, p = 0.0092). These differences remained significant overtime and at 24 hours WT cells completely close the scratch while Lhx2cKO‐derived cells presented only 39.7 % closure. Lhx2cKO corneal cells fully recovered at 48 hours after wounding (Fig. 4B).


LIM Homeobox Domain 2 Is Required for Corneal Epithelial Homeostasis.

Sartaj R, Chee RI, Yang J, Wan P, Liu A, Guaiquil V, Fuchs E, Rosenblatt MI - Stem Cells (2016)

Lhx2cKO derived corneal epithelial cells showed delayed wound healing.Corneal epithelial cells were isolated from WT and Lhx2cKO mice and cultured as indicated in Material and Methods. Cells were seeded in culture plates and when confluent a scratch was made through the cell monolayer and the wound closure followed via time‐lapse microscopy (A). Scratches in WT cells closed in less than 24 hrs, while closure of scratches made in Lhx2cKO cultures were significantly delayed, healing by 48hrs (B). Data are means SE ± (n = 3). *P ≤ 0.05. Abbreviation: WT, wild type.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4834794&req=5

stem2257-fig-0004: Lhx2cKO derived corneal epithelial cells showed delayed wound healing.Corneal epithelial cells were isolated from WT and Lhx2cKO mice and cultured as indicated in Material and Methods. Cells were seeded in culture plates and when confluent a scratch was made through the cell monolayer and the wound closure followed via time‐lapse microscopy (A). Scratches in WT cells closed in less than 24 hrs, while closure of scratches made in Lhx2cKO cultures were significantly delayed, healing by 48hrs (B). Data are means SE ± (n = 3). *P ≤ 0.05. Abbreviation: WT, wild type.
Mentions: Tissue injury is known to activate stem cells to proliferate and form new tissue in various adult organs 17, 18, 19, 20. In culture, we mimicked the corneal wound healing procedure by scratching confluent cultures of the WT and Lhx2cKO murine CECs and monitored the closure of the wound. Images were taken at 0, 8, 16, 24, and 48 hours at the same locations in the cultures using phase‐contrast microscopy. The cell‐free area of the scratch in WT and Lhx2cKO cells was measured at time 0 and selected for direct comparisons only if they had approximately the same wound area between replicas to monitor healing over time. We found a significant decrease in Lhx2cKO wound closure when compared with WT (Fig. 4A). Quantification of the cell‐free areas showed that Lhx2cKO cells delay wound healing as early as 8 hours (2.2 ± 1.16) when compared with the WT (7.5 ± 1.46, p = 0.0092). These differences remained significant overtime and at 24 hours WT cells completely close the scratch while Lhx2cKO‐derived cells presented only 39.7 % closure. Lhx2cKO corneal cells fully recovered at 48 hours after wounding (Fig. 4B).

Bottom Line: Immunodetection on corneal sections were used to visualize conjunctivalization, a sign of limbal barrier failure.Cell based assays showed that Lhx2cKO derived corneal epithelial cells have a significantly lower capacity to form colonies over time and delayed wound-healing recovery when compared to wildtype cells.We conclude that Lhx2 is required for maintenance of the corneal epithelial cell compartment and the limbal barrier.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Sciences, Illinois Eye and Ear Infirmary, University of Illinois at Chicago, Chicago, Illinois, USA.

No MeSH data available.


Related in: MedlinePlus