title: Primary culture of human Schwann and schwannoma cells: improved and simplified protocol.
abstract: Primary culture of human Schwann cells (SCs) and vestibular schwannoma (VS) cells are invaluable tools to investigate SC physiology and VS pathobiology, and to devise effective pharmacotherapies against VS, which are sorely needed. However, existing culture protocols, in aiming to create robust, pure cultures, employ methods that can lead to loss of biological characteristics of the original cells, potentially resulting in misleading biological findings. We have developed a minimally manipulative method to culture primary human SC and VS cells, without the use of selective mitogens, toxins, or time-consuming and potentially transformative laboratory techniques. Schwann cell purity was quantified longitudinally using S100 staining in SC cultures derived from the great auricular nerve and VS cultures followed for 7 and 12 weeks, respectively. SC cultures retained approximately ≥85% purity for 2 weeks. VS cultures retained approximately ≥80% purity for the majority of the span of 12 weeks, with maximal purity of 87% at 2 weeks. The VS cultures showed high level of biological similarity (68% on average) to their respective parent tumors, as assessed using a protein array featuring 41 growth factors and receptors. Apoptosis rate in vitro negatively correlated with tumor volume. Our results, obtained using a faster, simplified culturing method than previously utilized, indicate that highly pure, primary human SC and VS cultures can be established with minimal manipulation, reaching maximal purity at 2 weeks of culture. The VS cultures recapitulate the parent tumors' biology to a great degree, making them relevant models to investigate VS pathobiology.
authors.affiliation: Speech and Hearing Bioscience and Technology Program, Harvard - Massachusetts Institute of Technology, Division of Health Sciences and Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA; Eaton Peabody Laboratories and Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, 243 Charles Street, Boston, MA, 02114, USA.
authors.forename: Sonam
authors.initials: S
authors.lastname: Dilwali
authors.affiliation: Department of Otology and Laryngology, Harvard Medical School, 651 Huntington Avenue, Boston, MA 02115, USA; Eaton Peabody Laboratories and Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, 243 Charles Street, Boston, MA, 02114, USA.
authors.forename: Pratik B
authors.initials: PB
authors.lastname: Patel
authors.affiliation: Department of Otology and Laryngology, Harvard Medical School, 651 Huntington Avenue, Boston, MA 02115, USA; Eaton Peabody Laboratories and Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, 243 Charles Street, Boston, MA, 02114, USA.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Roberts
authors.affiliation: Department of Radiology, Massachusetts General Hospital, Boston, MA, USA.
authors.forename: Gina M
authors.initials: GM
authors.lastname: Basinsky
authors.affiliation: Department of Radiology, Massachusetts General Hospital, Boston, MA, USA.
authors.forename: Gordon J
authors.initials: GJ
authors.lastname: Harris
authors.affiliation: Department of Otology and Laryngology, Harvard Medical School, 651 Huntington Avenue, Boston, MA 02115, USA; Eaton Peabody Laboratories and Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, 243 Charles Street, Boston, MA, 02114, USA.
authors.forename: Kevin S
authors.initials: KS
authors.lastname: Emerick
authors.affiliation: Speech and Hearing Bioscience and Technology Program, Harvard - Massachusetts Institute of Technology, Division of Health Sciences and Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA; Department of Otology and Laryngology, Harvard Medical School, 651 Huntington Avenue, Boston, MA 02115, USA; Eaton Peabody Laboratories and Department of Otolaryngology, Massachusetts Eye and Ear Infirmary, 243 Charles Street, Boston, MA, 02114, USA. Electronic address: konstantina_stankovic@meei.harvard.edu.
authors.forename: Konstantina M
authors.initials: KM
authors.lastname: Stankovic
mesh_terms: D017209:Apoptosis; D049109:Cell Proliferation; D002478:Cells, Cultured; D006801:Humans; D066298:In Vitro Techniques; D009464:Neuroma, Acoustic / Q000473:pathology*; D061251:Primary Cell Culture / Q000379:methods*; D012583:Schwann Cells / Q000473:pathology*; D013997:Time Factors