Project description:We performed the next-generation sequencing for an isogenic WASP-KO macrophage model. Above 65 million clean reads per sample were obtained. We found the numerous RNA splicing asscoiated genes were overexpressed and thousands of mRNA events were aberrantly spliced in WASP-KO macrophages.
Project description:There is a significant demand for intermediate-scale bioreactors in academic and industrial institutions to produce cells for various applications in drug screening and/or cell therapy. However, the application of these bioreactors in cultivating hiPSC-derived immune cells and other blood cells is noticeably lacking. To address this gap, we have developed a xeno-free and chemically defined intermediate-scale bioreactor platform, which allows for the generation of standardized human iPSC-derived hematopoietic organoids and subsequent continuous production of macrophages (iPSC-Mac).
Project description:Macrophages are pivotal effectors of host immunity and regulators of tissue homeostasis. Understanding of human macrophage biology has been hampered by the lack of reliable and scalable models for cellular and genetic studies. Human iPSC-derived monocytes and macrophages, as an unlimited source of subject genotype-specific cells, will undoubtedly play an important role in advancing our understanding of the role of macrophage biology and implication in human diseases. Here, we present RNA-Seq data of hiPSC-derived monocytes and GM-CSF or M-CSF macrophages generated by a fully optimized differentiation protocol as well as primary myeloid cells.
Project description:Tissue resident macrophages are key players in inflammatory processes and their activation and functionality is crucial in health and disease. Diseases associated with alterations in homeostasis or dysregulation of the innate immune system are numerous and include allergic reactions, autoimmune diseases as well as cancer. Hence, these cells are of high interest in drug development. Currently, the main sources of macrophages used in drug development are still primary cells isolated from blood or tissue, or immortalized cell lines (e.g. THP-1). Here, we describe an improved method for large-scale production of tissue resident macrophages from induced pluripotent stem cells (iPSC) in unprecedented yields. For this, iPSC-derived macrophages are thoroughly characterized to confirm their cell identity and thus their suitability for screening purposes. We demonstrate that this method to generate macrophages from iPSC overcomes the limitations of using primary cells, i.e. donor variability and limited availability of large cell numbers. Notably, the cells recapitulate key functional characteristics, including cytokine release, phagocytosis and migration. Genetic modifications can be introduced at the macrophage progenitor stage, so this methodology will facilitate the generation of reporters and gain- and loss-of-function mutants in an isogenetic background, essential assets for target validation.
Project description:RNAseq was used to identify inflammation-related pathways activated in iPSC-derived hepatocytes by co-culture with iPSC-derived macrophages, thereby allowing for making inferences about cytokine mediators.
Project description:Resident cardiac macrophages are critical mediators of cardiac function. Despite their known importance to cardiac electrophysiology and tissue maintenance, there are currently no stem-cell derived models of human engineered cardiac tissues (hECT) that include resident macrophages. In this study, we made an iPSC-derived hECT model with a resident population of macrophages (iM0) to better recapitulate the native myocardium, and characterized their impact on tissue function. Macrophage retention within the hECTs was confirmed via immunofluorescence after 28 days of cultivation. Inclusion of iM0 significantly impacted hECT function, increasing contractile force production. A potential mechanism underlying these changes was revealed by interrogation of calcium signaling, which demonstrated modulation of β-adrenergic signaling in +iM0 hECTs. Collectively, these findings demonstrate that macrophages significantly enhance cardiac function in iPSC-derived hECT models, emphasizing the need to further explore their contributions not only in healthy hECT models, but also in the contexts of disease and injury.