title: [Purification and characterization of adult optic nerve head astrocytes].
abstract: OBJECTIVE
  To establish a method of purifying and characterizing adult astrocytes from optic nerve head (ONH).
  
  
  METHODS
  Experimental study. The lamina cribrosa tissue from ONH of human eye was isolated under anatomic microscopy, and then 4 to 6 little explants were incubated in each culture plate containing culture medium DMEM/F12. After 8 to 10 weeks, the cells were removed by digesting cells with 0.25% trypsogen. Selective astrocyte culture medium is subsequently used. After two passages, astrocytes were identified by the observation of cell morphology and immunofluorescent staining of GFAP and NCAM.
  
  
  RESULTS
  After 2 to 3 weeks of explants planting, cells showed an obvious migration procession by crawling in succession from the verge of the explants and rapidly splitting. Most cells displayed a flat star shape or polygon after digested with trypsogen. Several cells are long fusiformis. Almost all cells presented a flat star shape and simultaneously expressed GFAP and NCAM when the cells cultured with selective astrocyte culture medium.
  
  
  CONCLUSIONS
  Cultured human ONH astrocytes can be obtained by precisely separating lamina cribrosa and placing the explants on the margin of culture medium, a method that promotes cell adherence. Using selective astrocyte culture medium is very effective and convenient in purifying primary astrocytes.
authors.affiliation: Department of Ophthalmology, Fudan University, Shanghai, China.
authors.forename: Na
authors.initials: N
authors.lastname: Wu
authors.forename: Jian-jiang
authors.initials: JJ
authors.lastname: Xu
authors.forename: Sheng-hai
authors.initials: SH
authors.lastname: Zhang
authors.forename: Bo-qi
authors.initials: BQ
authors.lastname: Yang
authors.forename: Qi-hua
authors.initials: QH
authors.lastname: LE
authors.forename: Ji-hong
authors.initials: JH
authors.lastname: Wu
authors.forename: Xing-huai
authors.initials: XH
authors.lastname: Sun
mesh_terms: D000328:Adult; D001253:Astrocytes / Q000166:cytology*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D006801:Humans; D009898:Optic Disk / Q000166:cytology*