A Unified Methodological Framework for Vestibular Schwannoma Research

被引:7
|
作者
Landegger, Lukas D. [1 ,2 ,3 ]
Sagers, Jessica E. [1 ,4 ]
Dilwali, Sonam [1 ,4 ]
Fujita, Takeshi [1 ,2 ]
Sahin, Mehmet I. [1 ,2 ]
Stankovic, Konstantina M. [1 ,2 ,4 ]
机构
[1] Massachusetts Eye & Ear, Dept Otolaryngol, Eaton Peabody Labs, Boston, MA 02114 USA
[2] Harvard Med Sch, Dept Otolaryngol, Boston, MA 02115 USA
[3] Med Univ Vienna, Vienna Gen Hosp, Dept Otolaryngol, Vienna, Austria
[4] Harvard Med Sch, Program Speech & Hearing Biosci & Technol, Boston, MA 02115 USA
来源
关键词
Cancer Research; Issue; 124; Vestibular schwannoma; great auricular nerve; surgical sample; primary cell culture; tissue secretions; adeno-associated virus; perilymph sampling; Schwann cells; GROWTH; EPIDEMIOLOGY; CELLS; MANAGEMENT; EXPRESSION; THERAPY; VECTOR; TRENDS; DAMAGE; MIB-1;
D O I
10.3791/55827
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Vestibular schwannomas are the most common neoplasms of the cerebellopontine angle, making up 6-8% percent of all intracranial growths. Though these tumors cause sensorineural hearing loss in up to 95% of affected individuals, the molecular mechanisms underlying this hearing loss remain elusive. This article outlines the steps established in our laboratory to facilitate the collection and processing of various primary human tissue samples for downstream research applications integral to the study of vestibular schwannomas. Specifically, this work describes a unified methodological framework for the collection, processing, and culture of Schwann and schwannoma cells from surgical samples. This is integrated with parallel processing steps now considered essential for current research: the collection of tumor and nerve secretions, the preservation of RNA and the extraction of protein from collected tissues, the fixation of tissue for the preparation of sections, and the exposure of primary human cells to adeno-associated viruses for application to gene therapy. Additionally, this work highlights the translabyrinthine surgical approach to collect this tumor as a unique opportunity to obtain human sensory epithelium from the inner ear and perilymph. Tips to improve experimental quality are provided and common pitfalls highlighted.
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页数:9
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