3D in vitro test system for vestibular schwannoma

Analysis of ADAM9 Regulation and Function in Vestibular Schwannoma Primary Cells

“…Tissue samples and cell culture Tissue of 24 VS, surgically resected in the Department of Neurosurgery of the University Hospital Würzburg, was collected from January 2018 until July 2019. Half of each sample was embedded in para n for immunohistochemistry, the other half was processed for primary cell culture as described elsewhere [5]. Brie y, the fresh tumor samples were washed three times with ice-cold HBSS +/+ (Gibco, Carlsbad, CA, USA), shred into 0.2 × 0.2 cm small pieces and digested with 1.7 mg/ml dispase (PluriSTEM, Millipore, Massachusetts, USA) and 1.5 mg/ml collagenase (Sigma, Munich, Germany) in 6 ml VS-medium (DMEM supplemented with 10% heat-inactivated fetal bovine serum, 1% penicillin/streptomycin (all from Gibco, Carlsbad, CA, USA), 0.35% Glucose (B. Braun, Melsungen, Germany), 8 nM Heregulin (Stemcell Technologies, Cologne, Germany), 0.5 mM IBMX and 0.25 µg/ml insulin (both from Sigma, Munich, Germany)) for 17 h at 37°C and 5% CO 2 .…”

“…Brie y, the fresh tumor samples were washed three times with ice-cold HBSS +/+ (Gibco, Carlsbad, CA, USA), shred into 0.2 × 0.2 cm small pieces and digested with 1.7 mg/ml dispase (PluriSTEM, Millipore, Massachusetts, USA) and 1.5 mg/ml collagenase (Sigma, Munich, Germany) in 6 ml VS-medium (DMEM supplemented with 10% heat-inactivated fetal bovine serum, 1% penicillin/streptomycin (all from Gibco, Carlsbad, CA, USA), 0.35% Glucose (B. Braun, Melsungen, Germany), 8 nM Heregulin (Stemcell Technologies, Cologne, Germany), 0.5 mM IBMX and 0.25 µg/ml insulin (both from Sigma, Munich, Germany)) for 17 h at 37°C and 5% CO 2 . Afterwards, the suspension was centrifuged for 5 min at 300 ×g, the supernatant was discharged and the pellet resuspended in 12 ml VS-medium, before plating 2 ml into each well of laminated 6-well-plates [5]. The cells were cultured in a humidi ed atmosphere of 5% CO 2 at 37°C.…”

“…VS are slowly growing, benign tumors. Thus, it is a challenge to establish primary cell cultures [5] and even more di cult to achieve high transfection rates. For this reason, only a small number of experiments could be performed for protein knock down and overexpression, limiting generalizability of these experiments.…”

Analysis of ADAM9 regulation and function in vestibular schwannoma primary cells

“…Tissue of 24 VS, surgically resected in the Department of Neurosurgery of the University Hospital Würzburg, was collected from January 2018 until July 2019. Half of each sample was embedded in paraffin for immunohistochemistry, the other half was processed for primary cell culture as described elsewhere [6] and briefly outlined in Additional file 1.…”

“…VS are slowly growing, benign tumors. Thus, it is a challenge to establish primary cell cultures [6] and even more difficult to achieve high transfection rates. For this reason, only a small number of experiments could be performed for protein knock down and overexpression, limiting generalizability of these experiments.…”