Project description:Here we show that alveolar macrophage (AM) identity were conserved through culture expansion as determined by the analysis of epigenetic chromatin accessibility of core AM specific transcription markers and surface markers. Further, the adaptations to the culture environment were transient, reversible and fully restored to the in vivo status upon transplantation into the alveolar niche.
Project description:Naive, central memory (TCM), effector memory (TEM), and terminally differentiated effector memory RA (TEMRA, CD8+ only) T cell subsets were FACS separated from PBMC samples of four human donors using CCR7 and CD45RA as distinguishing cell surface markers. Samples were split and either immediately isolated, or incubated for 42-48 hours with anti-CD3/CD28 beads for ex-vivo stimulation and processed for ATAC-seq.
Project description:We report profiling of single cell chromatin accessbility and gene expression of human NK cells from CMV-seropositive and -negative healthy blood donors ex vivo, as well as chromatin remodeling after in vitro stimulation via NKG2C and/or IL-12+IL-18.
Project description:The aim was to assess miRNA expression in 3 human ex-vivo CD8+ T cell subsets which span from antigen inexperienced cells (Naïve) to early memory cells (central memory, Tcm) and later stage memory cells (effector memory, Tem) CD8+ T cells were sorted on a FACS Aria II machine. N = naïve = CD8+, CCR7+, CD45RA+, CD45RO-, Tcm = central memory = CD8+, CCR7+, CD45RA-, CD45RO-,Tem= effector memory = CD8+, CCR7-, CD45RA-, CD45RO+
Project description:Naive, central memory (TCM), effector memory (TEM), and terminally differentiated effector memory RA (TEMRA, CD8+ only) T cell subsets were FACS separated from PBMC samples of four human donors using CCR7 and CD45RA as distinguishing cell surface markers. Samples were split and either immediately isolated, or incubated for 42-48 hours with anti-CD3/CD28 beads for ex-vivo stimulation.
Project description:The aim was to assess miRNA expression in 3 human ex-vivo CD8+ T cell subsets which span from antigen inexperienced cells (NaM-CM-/ve) to early memory cells (central memory, Tcm) and later stage memory cells (effector memory, Tem) CD8+ T cells were sorted on a FACS Aria II machine. N = naM-CM-/ve = CD8+, CCR7+, CD45RA+, CD45RO-, Tcm = central memory = CD8+, CCR7+, CD45RA-, CD45RO-,Tem= effector memory = CD8+, CCR7-, CD45RA-, CD45RO+ PBMC were isolated from 3 healthy human donors and sorted by FACS into 3 CD8+ T cell subsets. Total RNA was purified using the miRVANA kit (Ambion)
Project description:We report profiling of single cell chromatin accessbility and gene expression of human NK cells from CMV-seropositive and -negative healthy blood donors ex vivo, as well as chromatin remodeling after in vitro stimulation via NKG2C and/or IL-12+IL-18.
Project description:Gene expression profiling of cells isolated ex vivo is a unique tool to assess gene expression in vivo. Exemplified for CD4+CD45RO+ effector/memory T helper (T E/M) lymphocytes of human peripheral blood, we have analyzed different isolation procedures and storage conditions for the introduction of bias.
Project description:Prospective, open labelled, multicenter trial to evaluate the feasibility of ex vivo culture 3D (chemogram obtaining) on biopsies in order to estimate the predictive value of this technique for treatment response in patients treated by two different chemotherapies (FOLFOX or FOLFIRI) for colorectal cancer.