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Jacquelyn M. Simonis, Hironori Uehara, YangKyung Cho, Subrata K. Das, Bonnie Archer, Romulo Albuquerque, Balamurali K. Ambati; p63+ Cells Can Be Isolated From Cadaver Corneas Up To 6 Weeks After Death. Invest. Ophthalmol. Vis. Sci. 2011;52(14):5127.
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To evaluate the proportion of stem cells derivable from cadaver corneas long after time of preservation. Currently, tissue for transplant is only suitable for a short period of time. Determining the duration of stem cell survival could indicate that donor corneas are viable for longer periods of time.
Human donor corneas were received from the Utah Lion’s Eye Bank. Limbal cells were isolated from the corneas by fine dissection and treatment with 0.25% trypsin-EDTA, and plated on a feeder layer of lethally irradiated 3T3-J2 cells (ATCC). One week later the cells were harvested and immunofluoresence was performed using antibodies for p63-a limbal stem cell marker and DAPI. Confocal microscopy was performed to evaluate the localization of p63 and DAPI positive cells.
Cells were successfully cultured from three donor corneas (M&F, age > 65) which had been harvested 6 weeks previously. Upon assessing p63 & DAPI co-localization it was found that approximately 10% of cultured cells were both p63 & DAPI positive.
Stem cells remain viable up to 6 weeks after preservation of donor cadaver corneas and can be successfully isolated, identified, and cultured in vitro.
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