sample 1
Homo sapiens
Protocols: Human brain microvascular endothelial cells (hBMECs) were cultured in 6-well plates then incubated either with live N. meningitidis or recombinant MafA or culture medium only (non-induced control) for 6 hours. Human BMEC (hBMEC/D3 cell line), was acquired from Merck/Millipore (Prague, Czech Republic). Cells were cultured in 6-well cell culture plates covered with collagen type I (Sigma, USA) and endothelial cell growth basal medium (EBM-2; Lonza,UK; Appendix 1). Cells were incubated at 37°C in a humid atmosphere in the presence of 5% CO2. N. meningitidis (NM, isolate M1/03, serogroup B, strain MC58).) was plated on commercially prepared Thayer Martin agar and a single isolated colony was transferred into 50 mL of Brain Heart Infusion Broth enriched with 10 mM MgCl2 . Bacteria were grown at 37°C and 5% CO2 until OD600= 0.6 (mid-exponential phase). Human brain endothelial cells cultured in 6-well plates were incubated either with live Neisseria meningitidis (MOI 0.5,bacterial cells were washed with minimal essential medium, Sigma) or recombinant MafA (27 μg/well). Nontreated cells were used as negative control, for 6 hours at 37°C in the presence of 5% CO2. mRNA from human brain microvascular endothelial cells was isolated using RNeasy Mini Kit (Qiagen, Germany) according to manufacturer's instructions. 250 ng of RNA were reverse transcribed with oligodT primers using QuantSeq 3′ mRNA-Seq Library Prep Kit (Lexogen, Austria).
ENA-FIRST-PUBLIC
2019-06-15T04:02:31Z
ENA-LAST-UPDATE
2019-05-30T17:04:53Z
External Id
SAMEA5669581
INSDC center name
Laboratory of Biomedical Microbiology and Immunology, The University of Veterinary medicine and Pharmacy in Kosice
INSDC first public
2019-06-15T04:02:31Z
INSDC last update
2019-05-30T17:04:53Z
INSDC status
public
Submitter Id
E-MTAB-8008:sample 1
broker name
ArrayExpress
cell line
hCMEC/D3
cell type
blood vessel endothelial cell
common name
human
organism part
brain
sample name
E-MTAB-8008:sample 1
scientific_name
Homo sapiens
stimulus
Neisseria meningitidis infection
time
6