Clinical application of induced pluripotent stem (iPS) cells is limited by low efficiency of iPS derivation, and protocols that permanently modify the genome to effect cellular reprogramming. Moreover, safe and effective means of directing the fate of patient-specific iPS cells towards clinically useful cell types are lacking. Here we describe a simple, non-mutagenic strategy for reprogramming cell fate based on administration of synthetic mRNA modified to overcome innate anti-viral responses. We show that this approach can reprogram multiple human cell types to pluripotency with efficiencies that greatly surpass established protocols. We further show that the same technology can be used to efficiently direct the differentiation of RNA-induced pluripotent stem (RiPS) cells into terminally differentiated myogenic cells. Our method represents a safe, efficient strategy for somatic cell reprogramming and directing cell fates that has broad applicability for basic research, disease modeling and regenerative medicine.
Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA.
Specimen part, Cell line
View SamplesHuman induced pluripotent stem (iPS) cells derived from somatic cells of patients hold great promise for modelling human diseases. Dermal fibroblasts are frequently used for reprogramming, but require an invasive skin biopsy and a prolonged period of expansion in cell culture prior to use. Here, we report the derivation of iPS cells from multiple human blood sources including peripheral blood mononuclear cells (PBMCs) harvested by routine venipuncture. Peripheral blood-derived human iPS lines are comparable to human embryonic stem (ES) cells with respect to morphology, expression of surface antigens, activation of endogenous pluripotency genes, DNA methylation and differentiation potential. Analysis of Immunoglobulin and T-cell receptor gene rearrangement revealed that some of the PBMC iPS cells were derived from T-cells, documenting derivation of iPS cells from terminally differentiated cell types. Importantly, peripheral blood cells can be isolated with minimal risk to the donor and can be obtained in sufficient numbers to enable reprogramming without the need for prolonged expansion in culture. Reprogramming from blood cells thus represents a fast, safe and efficient way of generating patient-specific iPS cells.
Reprogramming of T cells from human peripheral blood.
Specimen part, Cell line
View SamplesSmoking represents a major risk factor for chronic obstructive pulmonary disease (COPD), but it is difficult to characterize smoke-induced injury responses under physiological breathing conditions in humans. Here we generated small airway-on-a-chip microdevices lined by living human bronchiolar epithelium from normal or COPD patients and connected them to an instrument that 'breathes' whole cigarette smoke in and out of the chips to study smoke-induced pathophysiology in vitro. We used microarrays to detail the global program of gene expression in well-differentiated epithelial cells following smoke exposure to recapitulate clinical pathologies and identify disease-specific responses.
Matched-Comparative Modeling of Normal and Diseased Human Airway Responses Using a Microengineered Breathing Lung Chip.
Specimen part, Disease, Treatment
View SamplesRecent studies showing involvement of iNKT cells in lung viral infections and airway inflammation suggest that these cells are key players in both prevention and generation of immune-pathology in the lungs. It is not fully understood how iNKT cells are activated in the lungs and if this relied solely on lung dendritic cells. We recently showed that CD1d is expressed on airway epithelium, and now demonstrate that iNKT cells can be activated by primary airway epithelial cells, via both CD1d dependent and independent processes. Transcriptional analysis of human iNKT cells reveals that direct contact with lipid-pulsed primary human airway epithelial cells results in initiation of a programme of activation comprising rapid and concomitant induction of cytokine genes and genes to switch off this response. These findings establish a new mode of activation within the lungs for iNKT cells, and further enhance the role of airway epithelium in innate lung immunity.
No associated publication
Specimen part
View SamplesThis SuperSeries is composed of the SubSeries listed below.
Common genetic variants modulate pathogen-sensing responses in human dendritic cells.
Sex, Age, Race, Subject
View SamplesThis SuperSeries is composed of the SubSeries listed below.
The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity.
No sample metadata fields
View SamplesThe Cancer Cell Line Encyclopedia (CCLE) project is a collaboration between the Broad Institute, the Novartis Institutes for Biomedical Research and the Genomics Novartis Foundation to conduct a detailed genetic and pharmacologic characterization of a large panel of human cancer models
The Cancer Cell Line Encyclopedia enables predictive modelling of anticancer drug sensitivity.
No sample metadata fields
View SamplesThe NIH Roadmap Epigenomics Mapping Consortium aims to produce a public resource of epigenomic maps for stem cells and primary ex vivo tissues selected to represent the normal counterparts of tissues and organ systems frequently involved in human disease.
The NIH Roadmap Epigenomics Mapping Consortium.
Sex, Specimen part, Disease, Subject
View SamplesA reference collection of genome-wide transcriptional expression data for bioactive small molecules.
The Connectivity Map: using gene-expression signatures to connect small molecules, genes, and disease.
No sample metadata fields
View SamplesThis SuperSeries is composed of the SubSeries listed below.
Initial genome sequencing and analysis of multiple myeloma.
Specimen part, Disease, Disease stage
View Samples