title: GSM7111582: ChP, 7w, LV; Mus musculus; RNA-Seq
library_construction_protocol: Mice for all groups were first sedated as described above, transcardially perfused with PBS with 1% Heparin after which brain tissue was excised and lateral and fourth ventricle choroid plexus tissue were carefully microdissected and collected in serum-free RPMI at 4 °C. As choroid plexus tissue is small and does not allow for single mouse scRNA-Seq analyses, we pooled choroid plexus tissues from 8 mice to generate single pooled samples for naive 7-week-old (female), 22-week-old (male) and 82-week-old (male) mouse groups, for the lateral and fourth ventricle ChP tissue separately. After tissue collection, samples were pre-cut with small scissors and diluted and digested in 2:3 stock solution of CollagenaseI (10 U/ml, Worthington), Collagenase IV (400 U/ml, Worthington) and DNaseI (30 U/ml, Worthington). Incubation was at 37 °C for 30 min with up/down pipetting every 10 min. Samples were strained through a 70 μm filter (Falcon) and washed with a total of 700 μl RPMI. Samples were centrifuged for 7 min 300 g at 4 °C, supernatant discarded and pellet resuspended in 250 μl FACS buffer (1x HBSS without Ca 2+ /Mg 2+ , 2 mM EDTA, 2% BSA). Single live cells were sorted using the BD FACS ARIAII and loaded onto a Chromium GemCode Single Cell Instrument (10x Genomics) to generate single-cell gel beads in emulsion (GEMs). Libraries were prepared with the GemCode Single Cell 3' Gel Bead and Library Kit v1 (10x Genomics) and the Chromium i7 Multiplex Kit (10x Genomics) according to manufacturer's instructions. Afterwards,10X barcoded cDNA was prepared for Illumina HiSeq4000 Next Gen Sequencing. Sequencing was performed at the VIB Nucleomics Core (VIB, Leuven).