All authors of this manuscript have no conflicts of interest with regards to this research and its findings.
Original Research Article
An investigation of donor and culture parameters which influence epithelial outgrowths from cultured human cadaveric limbal explants†
Article first published online: 28 JAN 2013
Copyright © 2012 Wiley Periodicals, Inc.
Journal of Cellular Physiology
Volume 228, Issue 5, pages 1025–1030, May 2013
How to Cite
Baylis, O., Rooney, P., Figueiredo, F., Lako, M. and Ahmad, S. (2013), An investigation of donor and culture parameters which influence epithelial outgrowths from cultured human cadaveric limbal explants. J. Cell. Physiol., 228: 1025–1030. doi: 10.1002/jcp.24249
- Issue published online: 28 JAN 2013
- Article first published online: 28 JAN 2013
- Accepted manuscript online: 5 OCT 2012 07:49AM EST
- Manuscript Accepted: 26 SEP 2012
- Manuscript Received: 10 MAY 2012
Limbal stem cell deficiency is a blinding disease which affects the cornea at the front of the eye. The definitive cure involves replacing the corneal epithelial (limbal) stem cells, for example by transplanting cultured limbal epithelial cells. One method of performing cultures is to grow a sheet of epithelial cells from a limbal explant on human amniotic membrane. The growth of limbal tissue can be variable. The aim of this study is to investigate how different donor and culture factors influence the ex vivo growth of cadaveric limbal explants. Limbal explant cultures were established from 10 different cadaveric organ cultured corneo-scleral discs. The growth rate and the time taken for growth to be established were determined. Statistical analysis was performed to assess correlation between these factors and donor variables including donor age, sex, time from donor death to enucleation, time from enucleation to organ culture storage and duration in organ culture. Growth curves consistently showed a lag phase followed by a steeper linear growth phase. Donor age, time between death and enucleation, and time between enucleation and organ culture were not correlated to the lag time or the growth rate. Time in organ culture had a significant correlation with the duration of lag time (P = 0.003), but no relationship with the linear growth rate. This study shows that an important factor correlating with growth variation is the duration of corneo-scleral tissue in organ culture. Interestingly, donor age was not correlated with limbal explant growth. J. Cell. Physiol. © 2012 Wiley Periodicals, Inc.