Project description:Purpose: Our results show that in the absence of SLAMF5 ( by KO or blocking ab), there is an accumulation of Bregs (CD19+IL10+) and a specific increase of their survival. Here, using RNAseq, we wished to identify genes that take place in the regulation of Breg by SLAMF5 blocking. Method: Total RNA was extracted from 3x10e5 sorted CD19+GFP+cells from 2-3 Vert-x mice (induced with EAE) incubated with SLAMF5 blocking Ab or control IgG using the Direct-zol RNA kit (ZymoResearch) according to the manufacturer’s instructions. RNA-seq was performed using the MARS-seq protocol. Showing here 2 biological replicates of each group.
Project description:Human serum derived macrophages were infected with Mtb expressing a transcriptional reporter of viability and on day 5, the cells were sorted for RFP+GFP+ macrophages or RFP+GFP- macrophages.
Project description:CLDN5-GFP reporter has been engineered in hPSCs. These cells have been differentiated to Ecs. Differentaited Ecs were sorted into GFP+ and GFP- corresponding to CLDN5+ and CLDN5- to study gene expression of high-barrier resistance.
Project description:scRNA-seq on GFP + cells sorted from the Tg(mitfa:GFP) transgenic zebrafish embryos at 28 hours post fertilization (hpf) using the 10x Chromium system
Project description:To explore the potential function of CD19+ICOSL- B-cell-subset, the whole transcriptome of sorted CD19+ICOSL+ and CD19+ICOSL- B cells was profiled by using Microarray analysis.
Project description:Tissue engineering strategies that combine human pluripotent stem cell-derived myogenic progenitors (hPDMs) with advanced biomaterials provide promising tools for engineering 3D skeletal muscle grafts to model tissue development in vitro and promote muscle regeneration in vivo. We recently demonstrated (i) the potential for obtaining large numbers of hPDMs using a combination of two small molecules and (ii) the application of electrospun fibrin microfiber bundles for functional skeletal muscle restoration following volumetric muscle loss. In this study, we demonstrate that the biophysical cues provided by the microfiber bundles can induce unsorted hPDMs to form multinucleated myotubes that express desmin and myosin heavy chain. To reduce the batch-to-batch variability of these samples, we tested a genetic PAX7 reporter line (PAX7::GFP) to sort for more homogenous populations of hPDMs. RNA sequencing and gene set enrichment analyses confirmed that PAX7::GFP-sorted hPDMs exhibited higher expression of myogenic genes while unsorted hPDMs demonstrated increased expression of genes associated with embryonic, neural crest, and fibroadipogenic progenitor lineages. We tested engineered skeletal muscle grafts derived from either PAX7::GFP-sorted or unsorted hPDMs within in vivo skeletal muscle defects. The unsorted hPDM-derived grafts also exhibited greater fibrosis and more proinflammatory responses. In contrast, the PAX7::GFP-sorted groups had moderately high vascular infiltration, more implanted cell association with embryonic myosin heavy chain (eMHC) regions, and greater CD206:CD86 ratios compared to unsorted hPDMs and acellular fibers suggesting they induced more pro-regenerative microenvironments. These findings demonstrated some promise for the use of PAX7::GFP-sorted hPDMS on fibrin microfiber bundles and provided some insights for improving the cell-biomaterial system to stimulate more robust in vivo skeletal muscle regeneration.