Abstract
Purpose: :
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.
Methods: :
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.
Results: :
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.
Conclusions: :
Stem cells remain viable up to 6 weeks after preservation of donor cadaver corneas and can be successfully isolated, identified, and cultured in vitro.
Keywords: cell survival • cornea: storage • wound healing