GSM4119829: SUP (B-ALL/Niche/B-ALL-Niche); Homo sapiens; RNA-Seq
The leukemia BM niche samples (designated as B-ALL-Niche) were prepared in parallel with control groups of leukemia culture alone (designated as B-ALL alone) and niche cells (HUVEC, hMSCs, and hFOB) culture without leukemia (designated as Niche alone). The prepared leukemia models were cultured for 7 days in the media of a mixture (2:1:1:1) of HUVEC medium (EGMTM-2, Lonza), hMSCs medium (MSCGMTM, Lonza), hFOB medium (DMEM/F12) and Human B-ALL cell medium (RPMI1640 or IMDM, GIBCO). Single-cell suspensions of B-ALL samples were prepared by treating the ex vivo BM niche model devices with nattokinase (50 Fu/ml) for 45 min at 37 °C to release cells from the fibrin hydrogel, followed by centrifuging at 400×g at 4°C for 5 min. The cell pellets were re-suspended in PBS with 2% FCS and Human TruStain FcX Blocking Solution (BioLegend) and incubated on ice for 10 min. Cell suspensions were centrifuged at 400×g at 4°C for 5 min, and supernatant discarded. For cell hashing 6, cells retrieved from groups of B-ALL alone, Niche alone, and B-ALL-Niche were respectively labelled with three hashtag antibodies (TotalSeqTM, BioLegend), according to manufacturer's instructions. The mixed into two samples, REH and SUP samples. Single Cell 3' Reagent Kits: Chromium™ Single Cell 3' Library & Gel Bead Kit v3, PN-1000075; Single Cell 3' Chip Kit v3 PN-1000073 and i7 Multiplex Kit PN-120262 (10x Genomics). Samples were tagged with cell hashing oligo-tagged antibodies (BioLegend) according to manufacturer's instructions.
GEO Accession
GSM4119829