Project description:Human cerebral organoids (hCOs) offer the possibility of deepening the knowledge of human brain development, as well as the pathologies that affect it. The method developed here describes the efficient generation of hCOs by going directly from two-dimensional (2D) pluripotent stem cell (PSC) cultures to three-dimensional (3D) neuroepithelial tissue, avoiding dissociation and aggregation steps. This has been achieved by subjecting 2D cultures, from the beginning of the neural induction step, to dual-SMAD inhibition in combination with CHIR99021. This is a simple and reproducible protocol in which the hCOs generated develop properly presenting proliferative ventricular zones (VZs) formed by neural precursor and radial glia (RG) that differentiate to give rise to mature neurons and glial cells. The hCOs present additional cell types such as oligodendrocyte precursors, astrocytes, microglia-like cells, and endothelial-like cells. This new approach could help to overcome some of the existing limitations in the field of organoid biotechnology, facilitating its execution in any laboratory setting.
Project description:This is a simple and reproducible protocol in which the hCOs generated are homogeneous and develop properly presenting proliferative ventricular zones (VZs) formed by neural and radial glia (RG) precursors that differentiate to give rise to mature neurons and glia cells. The hCOs at four weeks of maturation present additional cell types such as oligodendrocyte precursors, astrocytes, microglia and endothelial cells as shown by single‐cell RNA sequencing (scRNA‐seq) and immunohistochemistry (IHC).
Project description:Transcriptional profile of PCSC spheres in SCM-1% KO (stem-like cells) vs adherent cultures in PCSC-Celprogen medium (differentiated-like cells) Two-condition experiment: Sphere vs. Parental/adherent cells. Biological replicates: 2 sphere replicates , 2 adherent replicates.
Project description:Transcriptional profile of LNCaP spheres in SCM-1% KO (stem-like cells) vs adherent cultures in RPMI-1640-10% FBS (differentiated-like cells)
Project description:To determine the effect of adherence on the metabolic and functional response of human monocytes. Monocytes were stimulated with lipopolysaccharide (LPS) under non-adherent and adherent conditions. To determine the role of glycolysis in LPS-induced immune responses, this pathway was inhibited by glucose deprivation or the glucose analog 2-deoxy-D-glucose (2DG).
Project description:MSC-adherent hematopoietic stem and progenotir cells (HSPC) express adhesion-associated genes at higher levels than non-adherent cells while cell-cycle and differentiation-associated genes are not significantly changed between the two cell populations. We used microarray to confirm identity of MSC-adherent and non-adherent cord blood-derived HSPCs and to exclude that cell cycle and differentiation affect adhesive capacity.
Project description:Transcriptional profile of PCSC spheres in SCM-1% KO (stem-like cells) vs adherent cultures in PCSC-Celprogen medium (differentiated-like cells) Two-condition experiment: Sphere vs. Parental/adherent cells. Biological replicates: 2 sphere replicates , 2 adherent replicates. Early passaged tumor cells were expanded in adherent conditions and then plated in sphere forming conditions (SCM-1KO% medium at low density) to isolate sphere growing cells.
Project description:Microarry based whole genome expression analysis was done for the more tumorigenic CaSki spheroids cultures and the less tumorigenic CaSki adherent cultures to identify signaling mechanisms that vary between the two populations. Subsequent to this, an independent array was done to characterize the differences in gene expression between the cancer stem cell-like Lin-CD66+ and the bulk Lin- CD66- cells isolated from primary human cervical carcinomas. The second array was custom-designed and had genes implicated in stemness, epithelial differentiation, signaling pathways implicated in stemness (Notch and Wnt) and cell cylce-regulation.