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Judith Zavala, Carlos-Alberto Rodriguez-Barrientos, Jorge E Valdez, ; Morphological profile analysis of cultured corneal endothelial cells in Mitogenic/Resting culture system for applications in tissue engineering.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5189.
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© ARVO (1962-2015); The Authors (2016-present)
To determine the benefits of including a resting culture medium after a mitogenic culture medium to maintenance hexagonal morphology in corneal endothelial cells.
4 young New Zealand white males rabbits were used in order to obtain corneo-escleral tissue. Corneal endothelial cells (CECs) were isolated. After five days in culture with mitogenic culture medium (mM), the first subculture was carried out and CECs was split into two populations; one of these were used to test the effect of the resting culture medium (rM) and the other one continued in culture with mM. After cell culture in P1 with mM and rM reached confluence (80-90%), cells were subculture (P2) and continued with the same scheme of culture medium. Morphological changes in mM and rM were photo-documented
Morphological changes in mM and rM became more evident after two days of incubation with each culture medium. Following for 14 days of culture, the effect of the mitogenic medium became more evident, morphology of CECs was severely altered, and cells showed more fibroblastic-like shape and form a high cell density monolayer.On the other hand, the cultured CECs in rM display important variations in cell morphology. Under phase contrast microscopy homogenous polygonal shape could be seen. CECs appear to be mostly polygonal with more tendency to hexagonality and homogenous during and throughout the five days of culture. The morphology was retained and cells grew well and form a dense coherent monolayer.
Using conventional techniques, CECs in culture present modification in their morphology. We support the use of a Mitogenic/Resting culture system for CECs. This step possesses the ability to maintain the hexagonallity morphology in cultured corneal endothelial cells.
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