In Situ
Hybridization of RA Induced P19 Cells
BIAN Wei, YANG Jing, DING Xiao-Yan# and JING
Nai-He *
( Shanghai Institute of Biochemistry, the Chinese Academy of Sciences,
Shanghai 200031, China;
# Shanghai Institute of Cell Biology, the Chinese Academy of
Sciences, Shanghai 200031, China )
Abstract P19 EC cells can be induced to
differentiate into neurons in vitro when they were treated with 1×10－6 mol/L retinoic acid. In the
process of neuronal differentiation, P19 cells gradually migrate and form
aggregates. Aggregated cells weakly adhered to the surface of cell culture
dishes are usually dropped of during the procedure of in situ hybridization. To
keep cells on the coverslips, we modified the procedure by using 1 g/L gelatin
solution to cover the cells and substituting proteinase K treatment with pepsin
treatment which digested cellular proteins mildly. With the treatment of
pepsin, RNA probe can easily go through cell membrane and bind to its target,
without loss of cell from coverslips.
Key Words P19 neuron; in situ
hybridization
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