title: Crystallin-β-b2-overexpressing NPCs support the survival of injured retinal ganglion cells and photoreceptors in rats.
abstract: OBJECTIVE
  Crystallin β-b2 (crybb2) is known to support the regeneration of retinal ganglion cell (RGC) axons in culture. We investigated whether neuronal progenitor cells (NPCs) overexpressing crybb2 (crybb2-NPC) affect secondary retinal degeneration due to optic nerve crush in vivo.
  
  
  METHODS
  NPCS were produced by dissociation and propagation of rat embryonic neural tube and eye primordial cells at embryonic days 13.5 and 15. Retinal degeneration was induced by injured optic nerve crush (BY suture, 20 seconds). Several groups were built: crybb2-NPC were injected into the vitreous body, while the Controls were comprised of recombinant crybb2-injected and PBS-injected groups. The eyes, in particular the retina, were analyzed by immunohistochemistry and Western blotting for different antigens at 2 and 4 weeks after surgery.
  
  
  RESULTS
  At 2 and 4 weeks post surgery, crybb2-NPC resided within the vitreoretinal compartment, and were persistently nestin-positive throughout the experimental period. The cells stained positive for various neurotrophins and acted as "living" cell factories to support the survival of injured RGCs. The crybb2-NPC migrated throughout the eye structures and sometimes became integrated within the tissue. Most of the ocular cells responded to the appearance of crybb2-NPC with marked changes of certain proteins, including Iba-1 (microglia), vimentin (glial cells), and rhodopsin (photoreceptors). Photoreceptors also displayed a better survival after crybb2-NPC injection compared to control groups.
  
  
  CONCLUSIONS
  Crybb2-NPC exert beneficial effects on the vitreoretinal compartment, which suggests that modified crybb2-NPC could be used in a novel strategy for the treatment of degenerative vitreoretinal diseases. However, future studies must determine the safety of in vivo administration of crybb2-NPC.
authors.affiliation: Institute for Experimental Ophthalmology, School of Medicine, Westfalian-Wilhelms-University Münster, Münster, Germany. Michael.boehm@ukmuenster.de
authors.forename: Michael R R
authors.initials: MR
authors.lastname: Böhm
authors.forename: Sarah
authors.initials: S
authors.lastname: Pfrommer
authors.forename: Carolin
authors.initials: C
authors.lastname: Chiwitt
authors.forename: Matthias
authors.initials: M
authors.lastname: Brückner
authors.forename: Harutyun
authors.initials: H
authors.lastname: Melkonyan
authors.forename: Solon
authors.initials: S
authors.lastname: Thanos
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002465:Cell Movement / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D004195:Disease Models, Animal; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D007150:Immunohistochemistry; D058449:Intravitreal Injections; D008297:Male; D009409:Nerve Crush; D009414:Nerve Growth Factors / Q000378:metabolism; D058953:Neural Stem Cells / Q000502:physiology*; D020419:Photoreceptor Cells, Vertebrate / Q000166:cytology* / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D012162:Retinal Degeneration / Q000378:metabolism / Q000517:prevention & control*; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000378:metabolism; D033581:Stem Cell Transplantation; D014162:Transfection; D038206:beta-Crystallin B Chain / Q000235:genetics*
chemical_list: D015415:Biomarkers; D009414:Nerve Growth Factors; D038206:beta-Crystallin B Chain; C113634:beta-crystallin B2