title: Human_Th0_72h_3
organism: Homo sapiens
description: Protocols: T cells from blood (human) and mouse (spleen) Mononuclear cells were isolated from the cord blood of healthy neonates at Turku University Central Hospital using Ficoll-Paque PLUS (GE Healthcare, #17-1440-02). CD4+ T cells were then isolated using the Dynal bead-based positive isolation kit (Invitrogen). CD4+ cells from three individual donors were activated directly in 24w plates with plate-bound anti-CD3 (500ng/well, Immunotech) and soluble anti-CD28 (500 ng/mL, Immunotech) at a density of 2×106 cells/mL of Yssel's medium67 containing 1% human AB serum (PAA). Th2 cell polarization was initiated with IL-4 (10 ng/ml, R&D Systems). Cells activated without IL-4 were also cultured (Th0). Total RNA was purified by Qiagen RNeasy Mini Kit according to manufacturer's instruction, and concentration was determined by a Nanodrop. A total of 500 ng RNA was used to prepare sequencing libraries using KAPA Stranded mRNA-seq Kit (KAPA #07962193001) according to manufacturer's instructions.
attributes.tag: ENA first public
attributes.value: 2017-12-12
attributes.tag: ENA last update
attributes.value: 2017-12-12
attributes.tag: External Id
attributes.value: SAMEA104423627
attributes.tag: INSDC center alias
attributes.value: Sanger Institute
attributes.tag: INSDC center name
attributes.value: Sanger Institute
attributes.tag: INSDC first public
attributes.value: 2017-12-12T17:01:54Z
attributes.tag: INSDC last update
attributes.value: 2017-12-12T11:42:18Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6300:Human_Th0_72h_3
attributes.tag: age
attributes.value: 0
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: T-helper 0 cell
attributes.tag: common name
attributes.value: human
attributes.tag: disease
attributes.value: normal
attributes.tag: individual
attributes.value: donor pool 3
attributes.tag: organism part
attributes.value: umbilical cord blood
attributes.tag: sample name
attributes.value: E-MTAB-6300:Human_Th0_72h_3
attributes.tag: stimulus
attributes.value: anti-CD28 (500 nanogram per milliliter)