characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2018-08-09T17:03:13Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2017-08-22T14:57:16Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA104223524
characteristics.characteristic: External Id
characteristics.text: Centre for Integrative Physiology, University of Edinburgh
characteristics.characteristic: INSDC center name
characteristics.text: 2018-08-09T17:03:13Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-08-22T14:57:16Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS1882542
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-5987:K4NAC
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: Rat microglia: to prepare rat microglia, on experimental day 1 10-12 wild-type Sprague-Dawley rat pups (mixed gender) of postnatal day 0-2 were decapitated. Their cortices were then dissected under a microscope and dissociated first chemically with papain solution and then mechanically with 5 ml pipettes. The cell suspension was then plated down in DMEM medium in poly-D-lysine coated T75 plates and grown at 37°C for 10-14 days. Human astrocytes: On day 2, primary human astrocytes (purchased from Caltag Medsystems, catalogue no. SC1800, which had been isolated from the cerebral cortex, and were negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi) were recovered from liquid nitrogen, plated down in human astrocyte medium first in a T75 flask and then split into 24-well plates the next day. They were then left to grow for 1-3 days until 80-90% confluency. Mouse neurons: On day 4 or 5, wild type CD1 mouse embryos (mixed gender) of day 17.5 were decapitated and cortices were dissected and dissociated as above and 9-13 X 104 cells (in NBA medium with 1% serum) per well were plated down on top of the human astrocytes prepared earlier. The neuron-astrocyte co-culture was left to grow for a further 3-4 days. On day 11 or 12, the rat microglia were harvested by shaking in an orbital shaker and plated down in NBA medium without serum on top of either neurons, astrocytes, or the neuron-astrocyte co-culture and were left to grow for 3 days. After 3 days of co-culture, the medium was replaced with Tmo for two hours. 500 ng LPS was then added to the cells for 12-16 hours. RNA was harvested using a Roche RNA isolation Kit according to standard protocol and the quality of total RNA (RIN) was confirmed on RNA 6000 Nano Chips and an Agilent 2100 Bioanalyzer following the manufacturer’s instructions. RNA-seq libraries were prepared by Edinburgh Genomics using the Illumina TruSeq stranded mRNA-seq kit, according to the manufacturer’s protocol (Illumina).
characteristics.characteristic: description
characteristics.text: co-culture between mouse neurons and human astrocytes
characteristics.characteristic: growth condition
characteristics.text: mixed sample
characteristics.characteristic: organism
characteristics.text: mixed sample
characteristics.characteristic: organism
characteristics.text: mixed sample
characteristics.characteristic: scientific_name
characteristics.text: K4NAC
characteristics.characteristic: title