2025-09-01
ENA first public
ERC000011
ENA-CHECKLIST
UOC
INSDC center name
public
INSDC status
ERS25250520
SRA accession
E-MTAB-15353:A006850315_207066_S108
Submitter Id
ArrayExpress
broker name
cd19 positive
cell type
not collected
collection date
human
common name
Protocols: Cocultures of CLL patient cells (n = 3) and primary human macrophages, THP-1 macrophages, NLCs, and mBMDM were set up as described in the manuscript. After 5 days of coculture, the medium of cocultures was collected, and containing CLL cells were purified using CD19 MicroBeads (Miltenyi Biotec), which magnetically label CD19+ cells and separate them via the magnetic field of a MACS Separator. The purification was performed using MACS LS Columns.  Primary and immortalized macrophage cells were used: mBMDM, NLC, primary human macrophages (HD-MDM), and THP-1 macrophages.  NLCs were generated by seeding patient-derived PBMCs and adherent cells were harvested after 14 days.  THP-1 monocytes were differentiated into macrophages via 100 ng/ml PMA for 48 h.   mBMDM were harvested from femurs and tibias of 8- to 12-week-old mice. Bones were flushed and cells were passed through a 100 μm sterile cell strainer. Erythrocytes were lysed with  ACK buffer and resulting cells cultured in DMEM (+ 1 % P/S, 10 % FBS) containing 20 ng/ml murine M-CSF  to pre-differentiate bone marrow stem cells into adherent M0 murine macrophages. After a minimum of 7 days, mBMDM were harvested.  HD-MDM were generated from buffy coats of healthy donors. After PBMCs isolation cells  were seeded in differentiation medium consisting of RPMI containing 10 % FBS, 1 % P/S, 50 ng/ml M CSF, 1 % NEAA, 1 % Na Pyruvate, 2 mM. Glutamax.  After 7 days, cells were collected.  For the isolation of RNA for Next Generation mRNA sequencing, the RNeasy Plus Mini Kit (Qiagen) was used according to the manufacturer’s instructions. Samples were processed and sequenced PE100, 35M reads per sample at the Cologne Center for Genomics according to in-house standards
description
NA
developmental stage
chronic lymphocytic leukemia
disease
not collected
geographic location (country and/or sea)
P182
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
NA
organism part
Homo sapiens
scientific_name
NA
sex
HD-MDM
stimulus
A006850315_207066_S108
title