Project description:Extracellular vesicles (EV) are secreted by nearly every mammalian cell type and contain a wealth of bioactive cargo capable of modulating target cell physiology and function though a variety of paracrine signaling mechanisms (Leavitt et al., 2019). Human embryonic stem cell (hESC)-derived extracellular vesicles (hESC-derived EV), were extracted from the hESC line H9 (WA09 Wicell Research Institute, Inc., Madison, WI). We analyzed the bioactive protein cargo to identify components that help to resolve radiation-induced injury to the lung in mice when injected in vivo.
Project description:In this study, we generated wildtype H9 hESC derived cardiomyocytes (CM) and neural stem cells (NSC) by in vitro differentiation. Global gene expression profiles were compared among undifferentiated H9 hESC and the derived CM and NSC. Comparison of global gene expression profiles of undifferentiated H9 hESC and the derived CM and NSC populations.
Project description:In this study, we generated wildtype H9 hESC derived cardiomyocytes (CM) and neural stem cells (NSC) by in vitro differentiation. Global gene expression profiles were compared among undifferentiated H9 hESC and the derived CM and NSC.
Project description:RNA-seq of H9 hESC line carrying shRB expression cassette, and H9 hESC line carrying shRB expression cassette in addition to CRISPR/Cas9-mediated nonsense mutations on RBL1 and RBL2
Project description:Small drug (SB/CHIR) treatment and orbital shear treatment promotes the generation of hemogenic endothelium and multipotent hematopoieitic progenitor cells. We used single cell RNA sequencing (scRNA-seq) to analyze the diversity of hemogenic niches derived from hESC H9 line at day 18 of in vitro differentiation.
Project description:We differentiated the human embryonic stem cell line H9 into retinal pigment epithelium (RPE) cells, to assess their transcriptomic profiles over time in culture. In-depth molecular analysis was performed by single cell RNA-Sequencing to access the molecular signature of RPE cells grown and harvested at two time points in culture (30 days and 369 days post passaging). We performed high-resolution comparisons at subpopulation and single-cell levels, to assess gene expression pathway signatures in RPE cells and upon aging in vitro. The hESC line H9 was grown to 70-80% confluence, transitioned to E6 medium for 2 days, with supplementation of N2 from day 2 until day 33. On day 33, medium was switched to RPEM (alpha-MEM, N1 supplement, 5% FBS, NEAA, Pen Strep Glutamine, taurine-hydrocortisone-triiodo-thyronin (THT)) for an additional 32 days. Cells were enzymatically passaged using 0.25% Trypsin EDTA and plated at 75,000 cells/cm2 on Matrigel growth factor reduced pre-coated plates (P1). Cells were subsequently harvested at day 30 (sample \\"YOUNG\\") and day 369 (sample \\"AGED\\"). Both samples were sorted for live cells using PI on a BD FACS Aria. Cells were centrifuged at 300g for 5 min and resuspended in PBS containing 0.04% BSA to a concentration of ~800-1000 cells/ µl. Approximately 17,400 cells were loaded onto a 10X chip single Cell 3′ Chips along with the reverse transcription master mix as per the manufacturer's protocol for the Chromium Single Cell 3′ v2 Library for a target recovery of 10,000 cells. Samples were then processed for sc Seq.
