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Differentiation Protocol-Dependent Variability in hiPSC-Derived Endothelial Progenitor Functionality

ABSTRACT: Differentiation Protocol-Dependent Variability in hiPSC-Derived Endothelial Progenitor Functionality

PROVIDER: PRJNA1165675 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30805715.fastq.gz	Fastqsanger.gz
	SRR30805716.fastq.gz	Fastqsanger.gz
	SRR30805717.fastq.gz	Fastqsanger.gz
	SRR30805718.fastq.gz	Fastqsanger.gz
	SRR30805719.fastq.gz	Fastqsanger.gz

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Differentiation Protocol-Dependent Variability in hiPSC-Derived Endothelial Progenitor Functionality

Project description:With multiple groups generating methods for differentiating induced pluripotent stem cells (hiPSCs) into endothelial cells/endothelial progenitors (ECs/EPs), there is a need to better understand the specific endothelial subtypes that different differentiation protocols produce. This is especially important for accurate tissue modeling as researchers continue to incorporate the endothelium into engineered tissues. We illustrated the heterogeneity of cells produced from different differentiation protocols by focusing on two selected protocols, one driving differentiation exclusively using small molecules (SM Protocol) and one driving differentiation primarily using growth factors (GF Protocol). We characterized the cells through a combination of vasculogenic computational analysis following encapsulation in 3D hydrogels, RNA sequencing, and measurement of soluble factor release. Vasculogenic computational analysis indicates that cells from the GF protocol formed more dense and interconnected vasculature compared to cells from the SM protocol. Likewise, RNA-seq analysis showed GF-derived cell enrichment in pathways involved in cell migration and angiogenesis. In addition, GF-derived cells favor differentiation to arterial endothelial cells as well as predisposition to undergoing a partial endothelial-to-mesenchymal transition, whereas SM-derived cells resemble immature progenitors based on higher proliferation rates as well as ECM remodeling. These trends also persisted following extended 3D culture. The results demonstrate that despite using the same starting hiPSC population and isolating EPs using the same surface marker, the two differentiation protocols yield highly distinct cell populations. This work highlights the importance of understanding the specific endothelial subtypes produced by different differentiation protocols for the creation of more accurate tissue models.

2024-09-26 | GSE278088 | GEO

Bulk RNA-seq of human induced pluripotent stem cells (hIPSC) and neural progenitor cells (NPC) differentiated usind Dual SMAD inhibition using the prime-seq method

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Development of cell therapy against CKD using functionally enhanced hiPSC-derived nephron progenitor cells (hiPSC-NPCs)

Project description:Development of cell therapy against CKD using functionally enhanced hiPSC-derived nephron progenitor cells (hiPSC-NPCs)

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Development of cell therapy against CKD using functionally enhanced hiPSC-derived nephron progenitor cells(hiPSC-NPCs)

Project description:Development of cell therapy against CKD using functionally enhanced hiPSC-derived nephron progenitor cells(hiPSC-NPCs)

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Total proteome analysis of HEK293, hiPSCs and hiPSC-derived neuronal progenitor cells, neurons and cardiomyocytes

Project description:DDA and DIA total proteome analysis of HEK293, kucg_2 hiPSC, and kucg_2 hiPSC-derived differentiated cells.

2025-05-02 | PXD044928 | Pride

Closing the mitochondrial permeability transition pore in hiPSC-derived endothelial cells induces glycocalyx formation and functional maturation.

Project description:Human induced pluripotent stem cells (hiPSCs) are used to study organogenesis and model disease as well as being developed for regenerative medicine. Endothelial cells are among the many cell types differentiated from hiPSC, but their maturation and stabilization fall short of that in adult endothelium. We examined whether shear stress alone or in combination with pericyte co-culture would induce flow alignment and maturation of hiPSC-derived endothelial cells (hiPSC-ECs) but found no effects comparable to those in primary microvascular EC. In addition, hiPSC-ECs lacked a luminal glycocalyx, critical for vasculature homeostasis, shear stress sensing and signalling. We noted however, that hiPSC-EC have dysfunctional mitochondrial permeability transition pores, resulting in reduced mitochondrial function and increased reactive oxygen species (ROS). Closure of these pores by cyclosporine-A improved EC mitochondrial function but also restored the glycocalyx such that alignment to flow took place. These indicated that mitochondrial maturation is required for proper hiPSC-EC functionality.

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Project description:The application potential of human induced pluripotent stem cells (hiPSC) derived human retinal organoids (HRO) relies on the robustness and transferability of the methodology for their generation. Standardized strategies and parameters to effectively assess, compare and optimize organoid protocols have been started to be established, but are not completed yet. To advance this, we explored the efficiency and reliability of a differentiation protocol that facilitates retina generation by formation of neuroepithelial cysts from hiPSC clusters. Here, we tested seven different hiPSC lines, which reproducibly generated HROs. Histologic and ultrastructural analyses support regulated HRO differentiation and maturation. The different hiPSC lines appeared to be a larger source of variance than experimental rounds. Whereas previous reports showed that HRO in several other protocols contain a rather low numbers of cones compared to rods, HROs derived by the cyst protocol consistently are cone-richer and with an comparable ratio of cones, rods, and Müller glia. Additionally, we devised a potential strategy to systematically evaluate different protocols side-by-side through parallel differentiation from the same hiPSC batches: The cyst-protocol was compared to a conceptually different protocol based on cell aggregate formation from single hiPSCs. Comparison of four hIPSC lines showed that both protocols reproduced key characteristics of retinal epithelial structure and cell composition, but the cyst-protocol provided a higher HRO yield. Further, while cyst-derived HROs maintained stable at least up to date 250, whereas single hiPSC-derived HROs showed spontaneous pathologic changes already by day 200. Overall, our data provide insight into the efficiency, reproducibility, and stability of the cyst-protocol for HRO generation, which will be useful for further organoid system optimization, as well as basic and translational research applications.

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Project description:The adult mammalian heart has been traditionally regarded as a post-mitotic organ with no regenerative capacity. However, this dogma has been refuted by some recent landmark studies. Both cardiac progenitor cells (CPCs) and epicardial progenitor cells (EPDCs) are activated after myocardium infarction and they may influence each other through paracrine mechanisms or direct interactions. Currently, efforts are being made to discover and develop therapeutic molecules to increase the number of CPCs and EPDCs in an infarcted heart. To better understand the characteristics and therapeutic potential of CPCs and EPDCs, as well as the regulatory mechanisms, we performed transcriptomic analysis of human iPSC-derived CPCs and human primary EPDCs and discovered unique gene expression profiles for each cell type. To make sure that the biological and pharmacological findings in cell assays under ambient oxygen conditions are relevant to the in vivo situation, it is important to understand the effect of hypoxia on the behavior, gene expression, and paracrine profiles of the cells. <br>Comparative transcriptomic analysis of human epicardial progenitor cells and hiPSC-derived cardiac progenitor cells was conducted. We performed global transcriptional analysis of two sources of cardiac progenitors, i.e., patient epicardium-derived cells (EPDCs) and cardiac progenitor cells (CPCs) derived from human induced pluripotent stem cells. In addition, we also compared the gene expression profiles of these cells when they were cultured under normoxic- and hypoxic conditions.

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Gene expression profilings of neural progenitor cell derived from hiPSC with ROR2 knockdown

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