SAMN32028213
External Id
Koltzov Institute of Developmental Biology of the Russian Academy of Sciences
INSDC center name
2022-12-04T00:00:00Z
INSDC first public
2022-12-04T15:20:01.855Z
INSDC last update
SRS15958427
INSDC secondary accession
live
INSDC status
Human
NCBI submission model
Human.1.0
NCBI submission package
SRS15958427
SRA accession
IDB RAS
biomaterial_provider
iMA
cell_line
EBS-protocol
cell_subtype
differentiating induced macrophages
cell_type
day -4 of differentiating
dev_stage
Homo sapiens
http://purl.obolibrary.org/obo/NCBITaxon_9606
organism
Human sample from Homo sapiens
title
The generation of iMacs using the EBS protocol (EBS-iMacs) was performed as described earlier [Nenasheva et al., 2020]. Briefly, iPSCs were expanded on MEFs, collected using collagenase IV in a way to preserve colony integrity, placed in ultralow adhesive 6-well plates and cultured in supplemented KO-DMEM medium (15% KOSR, 2µM Glutamax, 1% non-essential amino acids, 1% penicillin/streptomycin, 0,055mM β-mercaptoethanol) to allow EB formation (day -4). Four days later (day 0), the generated EBs were harvested, transferred to tissue-culture 6-well plates (15-20 EBs/well) and cultured in supplemented X-VIVO 15 medium (2µM Glutamax, 1% penicillin/streptomycin, 0,055mM β-mercaptoethanol) with the addition of IL-3 (25 ng/ml) and M-CSF (50 ng/ml). Full medium change was performed every 7 days. When floating monocytic precursors of iMacs appeared in the cultures, they were collected, transferred to new wells and cultured in supplemented RPMI-1640 medium (10% FCS, 2 µM L-Glutamax, 1% non-essential amino acids, 1 % penicillin/streptomycin, 0.055 mM β-mercaptoethanol) containing M-CSF (50 ng/ml) to induce terminal differentiation of EBS-iMacs. The remaining cultures were restimulated with supplemented X-VIVO 15 medium containing IL3/M-CSF for another round of iMac generation.
treatment