title: Neural stem cell-based intraocular administration of ciliary neurotrophic factor attenuates the loss of axotomized ganglion cells in adult mice.
abstract: OBJECTIVE
  To analyze the neuroprotective effect of intravitreally grafted neural stem (NS) cells genetically modified to secrete ciliary neurotrophic factor (CNTF) on intraorbitally lesioned retinal ganglion cells (RGCs) in adult mice.
  
  
  METHODS
  Adherently cultivated NS cells were genetically modified to express a secretable variant of mouse CNTF together with the fluorescent reporter protein Venus. Clonal CNTF-secreting NS cell lines were established using fluorescence activated cell sorting, and intravitreally grafted into adult mice 1 day after an intraorbital crush of the optic nerve. Brn-3a-positive RGCs were counted in flat-mounted retinas at different postlesion intervals to evaluate the neuroprotective effect of the CNTF-secreting NS cells on the axotomized RGCs. Anterograde axonal tracing experiments were performed to analyze the regrowth of the injured RGC axons in CNTF-treated retinas.
  
  
  RESULTS
  Intravitreally grafted NS cells preferentially differentiated into astrocytes that survived in the host eyes, stably expressed CNTF, and significantly attenuated the loss of the axotomized RGCs over a period of at least 4 months, the latest postlesion time point analyzed. Depending on the postlesion interval analyzed, the number of RGCs in eyes with grafted CNTF-secreting NS cells was 2.8-fold to 6.4-fold higher than in eyes with grafted control NS cells. The CNTF-secreting NS cells additionally induced long-distance regrowth of the lesioned RGC axons.
  
  
  CONCLUSIONS
  Genetically modified clonal NS cell lines may serve as a useful tool for preclinical studies aimed at evaluating the therapeutic potential of a sustained cell-based intravitreal administration of neuroprotective factors in mouse models of glaucoma.
authors.affiliation: Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Kai
authors.initials: K
authors.lastname: Flachsbarth
authors.affiliation: Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Kruszewski
authors.affiliation: Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Gila
authors.initials: G
authors.lastname: Jung
authors.affiliation: Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Wanda
authors.initials: W
authors.lastname: Jankowiak
authors.affiliation: Research Department Cell and Gene Therapy, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Riecken
authors.affiliation: Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Wagenfeld
authors.affiliation: Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Gisbert
authors.initials: G
authors.lastname: Richard
authors.affiliation: Research Department Cell and Gene Therapy, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Boris
authors.initials: B
authors.lastname: Fehse
authors.affiliation: Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Bartsch
mesh_terms: D000818:Animals; D002460:Cell Line; D020934:Ciliary Neurotrophic Factor / Q000008:administration & dosage*; D004195:Disease Models, Animal; D015316:Genetic Therapy / Q000379:methods*; D007150:Immunohistochemistry; D007267:Injections; D051379:Mice; D008810:Mice, Inbred C57BL; D058953:Neural Stem Cells; D009901:Optic Nerve Diseases / Q000473:pathology / Q000628:therapy*; D009915:Orbit; D012165:Retinal Ganglion Cells / Q000187:drug effects / Q000473:pathology*; D033581:Stem Cell Transplantation
chemical_list: D020934:Ciliary Neurotrophic Factor
keywords: axonal regeneration; ciliary neurotrophic factor; glaucoma; intraocular stem cell transplantation; lentiviral vector; neural stem cells; neuroprotection; optic nerve lesion; retinal ganglion cell