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Y. Liu, Z. C. Wang, P. Ma, Y. Xu, Z. P. Liu, A. P. Xiang; Embryonic Stem Cells and Their Conditioned Media Prevent Corneal Epithelial Cells From Aging. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5753.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the effects of the embryonic stem cells on anti-aging and dedifferentiation of mature corneal epithelial cells.
The corneal epithelial cells were cultured from rabbit peripheral corneal tissues, a region comprised of transient amplifying cells and less terminal differentiated cells, but without stem cells. The DMEM with high glucose were used in control group and experimental group 1 in co-culture with ES cells in Transwall systems, and the culture medium containing 30 % of the supernatant of conditioned media from culturing embryonic stem cells (E14 cell line) were used in experimental group 2. The ability of succession passage of the cells among three groups were compared; the morphology of corneal epithelial cells was observed under inverted phase contrast microscope; the cell growth curves were generated with MTT methods; the clone-forming ability was analyzed with Gimsa staining; the status of cell cycle was analyzed with flow cytometry.
1. Morphylogy: Corneal epithelial cells in the two experimental groups have apparent differences in cell morphology than those in control group, just like the cells of the P0 cells from the original tissue mass. 2. Cell growth curve: The time of P1 corneal epithelial cells entered the period of logarithmic growth phase were at day 3 for the cells in two experimental groups and at day 5 for the cells in the control group. 3. Clone-forming ability: Clone-forming ability of cells was 85 percent in both two experimental groups, this was significantly higher (P<0.05) than that of cells in the control group (about 40 percent). 4. Cell Cycle: Compared with the control group, the cells of the two experimental groups have a larger ratio in G1 phase (75.7% vs. 46.1%), and a lower ratio of cells in S phase (16.7% vs. 38.9%) of cell cycle.
Embryonic stem cells’ microenvironment could delay the aging of transient amplifying corneal epithelial cells, and may have a potential for supplying more progenitor cells for corneal tissue-engineering.
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