Polyethylene Terephthalate Membrane Used as a Holder for Cochlear Organotypic Cultures
LIU Yongze
ZHOU Han
MA Xiaofeng
MA Dengbin
GAO Xia
Abstract：Objective To report a practical method for tissue adhere in cochlear organotypic cultures. Methods The polyethylene terephthalate (PET) membrane was precoated with laminin, poly-L-ornithine, and fetal bovine serum at a ratio of 2:1:1 overnight. The cochlear basilar membrane was micro-dissected from C57BL/6J mouse pups at postnatal day 4, and placed on the PET membrane in a culture medium containing DMEM, 10%fetal bovine serum, and 30U/ml penicillin. After 48 hours, the cochlear specimens were fixed with 10%formalin in PBS. Anti-otoancorin and anti-tubulinⅢantibodies were dou?ble stained for labeling of nonsensory cells, interdental cells in the inner ear and auditory nerve fibers respectively. Fluores?cence conjugated phalloidin was also used to label stereocilia and cuticular plate of hair cells, and DAPI was also applied for nuclear staining. In addition, cochlear explants were also examined using scanning and transmission electron microscopy. Re?sults The cochlear basilar membrane firmly attached to the surface of PET. The microstructure and ultrastructure of cochlear cells appeared normal. Conclusion PET membrane can be used as an ideal holder for cochlear organotypic cultures.
Keywords：Polyethylene terephthalate membraneCochlear organotypic culturesHair cellInterdental cellImmunocy-tochemistryScanning electron microscopyTransmission electron microscopy
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 76-78 )