scRNAseq / snRNAseq of meningioma
A dissociation protocol was used to generate live cell populations from cryopreserved meningioma specimens. Samples were collected introperatively and immediately transferred and cryopreseerved. Samples were separately minced in dissociation media of 5ml of 0.1mg/ml Liberase TH, 1:1000 DNase I and gently rotated for 30 minutes. Dissociation was halted with a 5% FBS stop solution prior to resuspension in 1X RBC lysis buffer and viability assessment using a Countess. For tumour specimens that failed the dissociation protocol, nuclei isolation was performed by sectioning 100um curls on a cryostat without delay and collected in a 1.5ml centrifuge tube cooled on dry ice. Nuclei were isolated using the Chromium 10X nuclei isolation kit (CG000505 Rev A). Nuclei quality and quantity were assessed visually and using a Countess cell counter prior to FACS sorting usig 7-AAD and loading a Chromium Chip J. Libraries were generated in accordance with CG000338 Rev F Chromium Next GEM Single Cell Multiome ATAC + Gene Expression.