期刊
EXPERIMENTAL EYE RESEARCH
卷 137, 期 -, 页码 103-110出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.exer.2015.06.014
关键词
Astrocytes; Mouse; Optic nerve head; Piezo; Single-cell analysis; Single-cell isolation; TRPP
资金
- NIH [EY019703, EY022092]
- Lefler Foundation from Harvard Medical School
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). (C) 2015 Elsevier Ltd. All rights reserved.
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