Poster Presentation at 43rd Annual Meeting of The American Society of Cell Biology, San Francisco, California. 13-17 Dec 2003

Tissue regeneration for cornea using cord blood stem cells
N. Forraz,¹ S. Myriknas,² M. Baradez,¹ R. Ouvrard,¹ R. Lowe,¹ D. Swaby,² R. Morgan,¹ C. K. Rostron,² C. P. McGuckin¹ ; ¹ Basic Medical Sciences-Anatomy, Stem Cell Therapy Laboratory, London, United Kingdom, ² Basic Medical Sciences-Anatomy, St George's Hospital Medical School Keratec Eye Bank, London, United Kingdom
Presentation Number: L341
Poster Board Number: L341
Much debate surrounds embryological stem cell therapy. Less is known of the potential for stem cells from other sources. We have developed an umbilical cord blood (UCB) harvest strategy isolating ethically sound immature stem cell populations (UCBSC) with trans-differentiation potential (including neural, haemopoietic and hepatic populations). In the present work we investigated the capacity of these cells as vectors for tissue regeneration.
Success of corneal graft transplantation depends largely on the quality of the donor cornea, particularly the epithelium. In patients where this has been removed or destroyed, a potential exists for corneal ex vivo regeneration using explanted limbal corneal cells.
Donor-explanted limbal cell regions were cultured as superlayers on: (i) mitomicin-mitosis suppressed fibroblast feeder cells on amniotic membranes (AM) or ‘Laserskin®’ membrane; (ii) AM with cytokines; (iii) AM with cytokines and UCBSC. Set time point digital image capture measured growth aided by in-house Matlab developed software, (multiple spatial measurements (n=14-29) of distance between initial explant contours, to the expanded corneal border).
Basic explant growth was sustained but limited by the fibroblasts lifespan. On ‘Laserskin’ base, limbal explants demonstrated expansion of 0.10mm (SD=± 0.04) after 6 days and up to 2.07mm (SD=± 0.52) after 22 days [expansion rate 0.01mm/day at day 0 to 6 with exponential increase to an optimal rate of 0.12mm/day between day 6 and 22].
Overall, limbal explants achieved > 164% growth in 22 days. Upon UCBSC / cytokine supplementation (including FGF-1 and VEGF) the fibroblast feeding effect could be replaced with wave outcrops of developing corneal cells across the substrata being observed.
To investigate if UCBSC or cytokines caused the effect, porous culture inserts were inserted between the limbal corneal grafts and UCBSC. Growth was achieved, but severely slowed suggesting a direct cell to cell contact role for the UCBSC in the limbal corneal regeneration.
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