Project description:In vitro cardiac differentiation of human ESCs recapitulates in vivo embryonic heart development, and thus, serves as an excellent tool to investigate human cardiac development. Identification of molecular signatures during cardiac differentiation of human ESCs is instrumental for advancing our understanding of human cardiogenesis. We, as well as others have improved cardiac differentiation protocols significantly in recent years; however, detailed molecular mechanisms involved in cardiac lineage commitment have not yet been clearly defined. Based on this, we tried to identify the cellular hierarchies and molecular signatures of each of the in vitro human ESC-differentiating cardiac cell lineages through the well-established cardiac differentiatio protocol by Wnt signaling modulation and the FACS-sorted population RNA-seq analyses.
Project description:Gata5 efficiently promotes cardiomyocyte fate from murine ESCs.By removing serum from the culture conditions, GATA4 and GATA6 are each also able to efficiently promote cardiogenesis in ESC derivatives, with some distinctions. Thus, GATA factors can function in ESC derivatives upstream of other cardiac transcription factors to direct specification of progenitors with cardiac potential. We used microarray to compared the global gene expression of Gata5-induced cardiac cells with other growth factor directed ESC-derived cardiac cells
Project description:Gata5 efficiently promotes cardiomyocyte fate from murine ESCs.By removing serum from the culture conditions, GATA4 and GATA6 are each also able to efficiently promote cardiogenesis in ESC derivatives, with some distinctions. Thus, GATA factors can function in ESC derivatives upstream of other cardiac transcription factors to direct specification of progenitors with cardiac potential. We used microarray to compared the global gene expression of Gata5-induced cardiac cells with other growth factor directed ESC-derived cardiac cells iGata5 ESCs were induced with doxycycline at day 4 and harvested at day16 for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Long noncoding RNAs (lncRNAs) have emerged as crucial regulators of gene expression during embryonic stem cell (ESC) self-renewal and differentiation. Here, we systemically analyzed the differentially regulated lncRNAs during ESC-derived cardiomyocyte (CM) differentiation. We established a perspicuous profile of lncRNA expression at four critical developmental stages and found that the differentially expressed lncRNAs were grouped into six distinct clusters. The cluster with specific expression in ESC enriches the largest number of lncRNAs. Investigation of lncRNA-protein interaction network revealed that they are not only controlled by classic key transcription factors, but also modulated by epigenetic and epitranscriptomic factors including N6-methyladenosine (m6A) effector machineries.
Project description:We studied the effects of different stimuli on on the subcellular proteome of neurons differentiated from murine embryonic stem cells (ESC). We focused on the changes in the chromatin-bound fraction to elucidate specific differences in the gene regulatory machinery activated upon stimulation with Brain-derived neurotrophic factor (BDNF) or a general membrane-depolarization stimulus, potassium chloride (KCl).
Project description:The identification of cell surface proteins on stem cells or stem cell derivatives is a key strategy for the functional characterization, isolation, and understanding of stem cell population dynamics. Here, using an integrated mass spectrometry and microarray based approach, we analyzed the surface proteome and transcriptome of cardiac progenitor cells (CPCs) generated from the stage-specific differentiation of mouse and human pluripotent stem cells. Through bioinformatic analysis, we have identified and characterized FZD4 as a new marker for lateral plate mesoderm. Additionally, we utilized FZD4, in conjunction with FLK1 and PDGFRA, to further purify CPCs and increase cardiomyocyte (CM) enrichment in both mouse and human systems. Moreover, we have shown that NORRIN presented to FZD4 further increases CM output via proliferation through the canonical WNT pathway. Taken together, these findings demonstrate a role for FZD4 in mammalian cardiac development.
Project description:A hESC MESP1 reporter line was used to isolate MESP1 expressing pre-cardiac progenitors. These progenitor were replated and fulter differentiated in culture. At four sequential timepoints upon further differentiation, samples were isolated for gene expression analysis, in order to identify key cardiac transcription factors and molecules. hESCs were differentiated towards the cardiac lineage. MESP1-mCherry expressing progenitors were isolated at day 3 of differentiation and replated as aggregates, in presence of Wnt-inhibitor Xav939. Two days after replating (D5), four days (D7), seven days (D10), ad 11 days (D14), total RNA of each sample was isolated for gene expression analysis. MESP1-mCherry positive derivatives were compared to MESP1-mCherry negative derivatives, in order to identify cardiac-specific regulators and cell surface markers.
Project description:The goal of this experiment was to determine whether transcriptionally similar ESC-derived and primary donor cells give rise to similar donor-derived cells following transplantation into bleomycin injured lungs.