Using sterile technique, freshly harvested human VS tissue was rinsed in phosphate buffered saline (PBS) and dissected in culture medium consisting of Dulbecco’s Modified Eagle’s medium with Ham’s F12 mixture (DMEM/F12), 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 1% GlutaMAX. VS tissue was then dissociated in hyaluronidase and collagenase (all Life Technologies, Carlsbad, CA, USA) overnight, and cultured for 2–4 weeks, as described previously32 (link). Informed consent was obtained from all patients. The study protocols were approved by the Human Studies Committee of Massachusetts General Hospital and Massachusetts Eye and Ear Infirmary, and conducted in accordance with the Helsinki Declaration.
In addition to a primary human VS cell culture, two cell lines were used: 1) HEI-193 cells, an immortalized cell line derived from the vestibular schwannoma of a human NF2 patient, and 2) Nf2−/− SC4 cells, a mouse Schwann cell line (both gifts from the House Ear Institute, Los Angeles, CA, USA). The cell lines were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin.
In addition to a primary human VS cell culture, two cell lines were used: 1) HEI-193 cells, an immortalized cell line derived from the vestibular schwannoma of a human NF2 patient, and 2) Nf2−/− SC4 cells, a mouse Schwann cell line (both gifts from the House Ear Institute, Los Angeles, CA, USA). The cell lines were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin.
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