Increased integration of transplanted CD73-positive photoreceptor precursors into adult mouse retina.
Invest Ophthalmol Vis Sci, 2011/8/16;52(9):6462-71.
Eberle D[1], Schubert S, Postel K, Corbeil D, Ader M
Affiliations
PMID: 21743009DOI: 10.1167/iovs.11-7399
Impact factor: 4.925
Abstract
PURPOSE. Retinal degeneration initiated by loss of photoreceptors is the prevalent cause of visual impairment and blindness in industrialized countries. Transplantation of photoreceptor cells represents a possible replacement strategy. This study determined that identification of cell surface antigens can assist in enriching photoreceptor precursors for transplantation. METHODS. The expression profile of rod photoreceptors at postnatal day 4 was investigated by microarray analysis to identify photoreceptor-specific cell surface antigens. For enrichment of transplantable photoreceptors, neonatal retinas from rod photoreceptor-specific reporter mice were dissociated, and the rods were purified by magnetic associated cell sorting (MACS) with CD73 antibodies. MAC-sorted cell fractions were transplanted into the subretinal space of adult wild-type mice. The number of rod photoreceptors contained in unsorted, CD73-negative, and CD73-positive cell fractions were quantified in vitro and after grafting in vivo. RESULTS. Microarray analysis revealed that CD73 is a marker for rod photoreceptors. CD73-based MACS resulted in enrichment of rods to 87%. Furthermore, in comparison with unsorted cell fractions, CD73-positive MAC-sorted cells showed an approximately three-fold increase in the number of integrated, outer segment-forming photoreceptors after transplantation. CONCLUSIONS. CD73-based MACS is a reliable method for the enrichment of integrating photoreceptors. Purification via cell surface markers represents a new tool for the separation of transplantable photoreceptor precursors from a heterogeneous cell population, avoiding the need of reporter gene expression in target cells.
MeSH terms
5'-Nucleotidase; Animals; Animals, Newborn; Basic-Leucine Zipper Transcription Factors; Biomarkers; Cell Culture Techniques; Cell Lineage; Cell Tracking; Cell Transplantation; Eye Proteins; Flow Cytometry; Fluorescent Antibody Technique, Indirect; Gene Expression Profiling; Green Fluorescent Proteins; Immunomagnetic Separation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Oligonucleotide Array Sequence Analysis; Polymerase Chain Reaction; Retinal Detachment; Retinal Rod Photoreceptor Cells; Rhodopsin; Stem Cell Transplantation
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