title: Isolation of intact astrocytes from the optic nerve head of adult mice.
abstract: The astrocytes of the optic nerve head are a specialized subtype of white matter astrocytes that form the direct cellular environment of the unmyelinated ganglion cell axons. Due to their potential involvement in glaucoma, these astrocytes have become a target of research. Due to the heterogeneity of the optic nerve tissue, which also contains other cell types, in some cases it may be desirable to conduct gene expression studies on small numbers of well-characterized astrocytes or even individual cells. Here, we describe a simple method to isolate individual astrocytes. This method permits obtaining astrocytes with intact morphology from the adult mouse optic nerve and reduces contamination of the isolated astrocytes by other cell types. Individual astrocytes can be recognized by their morphology and collected under microscopic control. The whole procedure can be completed in 2-3 h. We also discuss downstream applications like multiplex single-cell PCR and quantitative PCR (qPCR).
authors.affiliation: Department of Ophthalmology, Harvard Medical School, Massachusetts Eye and Ear Infirmary, Boston, United States.
authors.forename: Hee Joo
authors.initials: HJ
authors.lastname: Choi
authors.affiliation: Department of Ophthalmology, Harvard Medical School, Massachusetts Eye and Ear Infirmary, Boston, United States.
authors.forename: Daniel
authors.initials: D
authors.lastname: Sun
authors.affiliation: Department of Ophthalmology, Harvard Medical School, Massachusetts Eye and Ear Infirmary, Boston, United States. Electronic address: Tatjana_Jakobs@meei.harvard.edu.
authors.forename: Tatjana C
authors.initials: TC
authors.lastname: Jakobs
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000473:pathology*; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002478:Cells, Cultured; D005901:Glaucoma / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009898:Optic Disk / Q000473:pathology*
keywords: Astrocytes; Mouse; Optic nerve head; Piezo; Single-cell analysis; Single-cell isolation; TRPP