Abstract
Introduction: :
2010 ARVO Abstract G. Qiu Dec. 2009Tina Guanting Qiu1**, Debra A Carter1, Dave Copland1, Andrew A Dick1Bristol Eye Hospital, University of Bristol, Bristol BS1 2LX United KingdomLower Maudlin Street, Bristol, BS1 2LX, UK
Purpose: :
To introduce a unique concept of photoreceptor reprogramming to generate high profile human photoreceptor lineage progenitors for retinal cell therapy.Materials &
Methods: :
Sixty-four pairs of post-mortem human eyes were obtained from Bristol Eye Bank with research consent and ethical approval. A series of cell isolation and stage specific purification tactics were developed to trigger the cell reprogramming in vitro, and a unique stepwise in vitro cell manipulations in combination with serum free define culture media were developed and validated. The evolving cell behaviours were observed and documented by light microscopy. Phenotypic and genotypic analysis was performed.
Results: :
The study first demonstrated that post-mitotic human photoreceptors be reprogrammed toward a precursor stage expressing nestin and a proliferation marker (Ki-67), and forming a robust neuronal network in vitro. qPCR confirmed the nestin up-regulation in the reprogrammed cell population. The majority of the cultured cells expressed photoreceptor cell markers (recoverin or rhodopsin positive) with negligible glial lineage phenotypes. High resolution 3D confocal imaging demonstrated a unique "tree-like" or "grape-like" structure of the cultured photoreceptor cells centring around the neurosphere.
Conclusions: :
The unique concept of cell reprogramming (turn the clock back at a certain limit) opens an alternative therapeutic pathway for regenerative medicine. The ability to generate high profile human photoreceptor lineage progenitor cell from adult tissue provides a valuable human cell source for photoreceptor replacement.Acknowledgements: 1. National Eye Research Centre, UK (Financial Support); 2. Bristol Tissue Eye Bank, UK; 3.Research Enterprise Development, University of Bristol, UK.Patent (Pending): UK Application No. 0913109.5 **Inventor:Guanting Qiu.** Corresponding author: Guanting Qiu, MD PhD, University of Bristol, Email: guantingqiu@yahoo.comPatent Filing No.: UK Application no. 0913109.5 (Pending)
Keywords: retina • photoreceptors • development