title: Increased integration of transplanted CD73-positive photoreceptor precursors into adult mouse retina.
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.
authors: DFG-Center for Regenerative Therapies Dresden (CRTD), Cluster of Excellence, Dresden, Germany.
authors: Dominic
authors: D
authors: Eberle
authors: Sandra
authors: S
authors: Schubert
authors: Kai
authors: K
authors: Postel
authors: Denis
authors: D
authors: Corbeil
authors: Marius
authors: M
authors: Ader
mesh_terms: D015720:5'-Nucleotidase / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D050976:Basic-Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism*; D018929:Cell Culture Techniques; D019070:Cell Lineage; D058948:Cell Tracking; D017690:Cell Transplantation; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D018189:Immunomagnetic Separation; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D012163:Retinal Detachment / Q000201:enzymology / Q000473:pathology / Q000628:therapy*; D017948:Retinal Rod Photoreceptor Cells / Q000166:cytology* / Q000201:enzymology; D012243:Rhodopsin / Q000235:genetics / Q000378:metabolism; D033581:Stem Cell Transplantation
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; D015415:Biomarkers; D005136:Eye Proteins; C084884:Nrl protein, mouse; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D012243:Rhodopsin; D015720:5'-Nucleotidase