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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 had a reduced capacity to form colonies in vitro freshly isolated WT and Lhx2cKO corneal epithelial cells were seeded in six‐well plates and allowed to form colonies as indicated in Material and Methods section. Phase contrast and crystal violet staining showed that Lhx2cKO‐derived cells have a reduced capacity to form colonies when compared with WT cells (A). Initially Lhx2cKO‐derived cells formed more colonies at day 5 in culture, after day 9, they only reached 50% of those formed by WT‐derived cells. Quantification of the colony forming efficiency on crystal violet staining showed a significant decrease in Lhx2cKO compared with WT mice (B). Scale bar =  200 µM. Data are means ± SE (n = 3). *, p  ≤ 0.05. Abbreviations: CFE, colony forming efficiency; WT, wild type.
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stem2257-fig-0003: Lhx2cKO‐derived corneal epithelial cells had a reduced capacity to form colonies in vitro freshly isolated WT and Lhx2cKO corneal epithelial cells were seeded in six‐well plates and allowed to form colonies as indicated in Material and Methods section. Phase contrast and crystal violet staining showed that Lhx2cKO‐derived cells have a reduced capacity to form colonies when compared with WT cells (A). Initially Lhx2cKO‐derived cells formed more colonies at day 5 in culture, after day 9, they only reached 50% of those formed by WT‐derived cells. Quantification of the colony forming efficiency on crystal violet staining showed a significant decrease in Lhx2cKO compared with WT mice (B). Scale bar =  200 µM. Data are means ± SE (n = 3). *, p  ≤ 0.05. Abbreviations: CFE, colony forming efficiency; WT, wild type.

Mentions: We assessed the proliferative capacity, population doublings, and colony forming efficiency (CFE) of WT versus Lhx2cKO corneal epithelial cells (CECs) in culture. Cells were isolated from the entire cornea and evaluated in a time dependent manner for CFE, and at day 12, the cultures were stained with crystal violet to visualize the colonies. In general, Lhx2cKO‐derived cells showed reduced cell proliferation and CFE efficiency when compared with WT cells (Fig. 3A). Initially at day 5, Lhx2cKO CECs showed increased CFE (calculated by the number of colonies divided by the total number of plated cells on a 3T3 feeder layer) by 37% (CFE WT = 1 ± 0.05% vs. Lhx2cKO = 1.37 ± 0.06%, p = 0.0027) compared with the WT cells. The trend changed after day 7 and WT cells were 30.5% higher than Lhx2cKO cells (CFE WT =  2.3 ± 0.2% vs. Lhx2cKO = 1.6 ± 0.1%, p = 0.006) and at day 9, CFE were 50% higher (CFE WT = 4.1 ± 0.30% vs. Lhx2cKO = 2.1 ± 0.2%, p = 0.001) (Fig. 3B). Lhx2cKO corneal epithelial cells were able to form colonies in culture, though greater numbers of colonies were predominantly seen in WT cultures and rarely identified in the Lhx2 deficient cells.


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 had a reduced capacity to form colonies in vitro freshly isolated WT and Lhx2cKO corneal epithelial cells were seeded in six‐well plates and allowed to form colonies as indicated in Material and Methods section. Phase contrast and crystal violet staining showed that Lhx2cKO‐derived cells have a reduced capacity to form colonies when compared with WT cells (A). Initially Lhx2cKO‐derived cells formed more colonies at day 5 in culture, after day 9, they only reached 50% of those formed by WT‐derived cells. Quantification of the colony forming efficiency on crystal violet staining showed a significant decrease in Lhx2cKO compared with WT mice (B). Scale bar =  200 µM. Data are means ± SE (n = 3). *, p  ≤ 0.05. Abbreviations: CFE, colony forming efficiency; WT, wild type.
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stem2257-fig-0003: Lhx2cKO‐derived corneal epithelial cells had a reduced capacity to form colonies in vitro freshly isolated WT and Lhx2cKO corneal epithelial cells were seeded in six‐well plates and allowed to form colonies as indicated in Material and Methods section. Phase contrast and crystal violet staining showed that Lhx2cKO‐derived cells have a reduced capacity to form colonies when compared with WT cells (A). Initially Lhx2cKO‐derived cells formed more colonies at day 5 in culture, after day 9, they only reached 50% of those formed by WT‐derived cells. Quantification of the colony forming efficiency on crystal violet staining showed a significant decrease in Lhx2cKO compared with WT mice (B). Scale bar =  200 µM. Data are means ± SE (n = 3). *, p  ≤ 0.05. Abbreviations: CFE, colony forming efficiency; WT, wild type.
Mentions: We assessed the proliferative capacity, population doublings, and colony forming efficiency (CFE) of WT versus Lhx2cKO corneal epithelial cells (CECs) in culture. Cells were isolated from the entire cornea and evaluated in a time dependent manner for CFE, and at day 12, the cultures were stained with crystal violet to visualize the colonies. In general, Lhx2cKO‐derived cells showed reduced cell proliferation and CFE efficiency when compared with WT cells (Fig. 3A). Initially at day 5, Lhx2cKO CECs showed increased CFE (calculated by the number of colonies divided by the total number of plated cells on a 3T3 feeder layer) by 37% (CFE WT = 1 ± 0.05% vs. Lhx2cKO = 1.37 ± 0.06%, p = 0.0027) compared with the WT cells. The trend changed after day 7 and WT cells were 30.5% higher than Lhx2cKO cells (CFE WT =  2.3 ± 0.2% vs. Lhx2cKO = 1.6 ± 0.1%, p = 0.006) and at day 9, CFE were 50% higher (CFE WT = 4.1 ± 0.30% vs. Lhx2cKO = 2.1 ± 0.2%, p = 0.001) (Fig. 3B). Lhx2cKO corneal epithelial cells were able to form colonies in culture, though greater numbers of colonies were predominantly seen in WT cultures and rarely identified in the Lhx2 deficient cells.

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