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Stem Cell Reports Oct 2023The formation of vascular structures is fundamental for in vitro tissue engineering. Vascularization can enable the nutrient supply within larger structures and...
The formation of vascular structures is fundamental for in vitro tissue engineering. Vascularization can enable the nutrient supply within larger structures and increase transplantation efficiency. We differentiated human induced pluripotent stem cells toward endothelial cells in 3D suspension culture. To investigate in vitro neovascularization and various 3D microenvironmental approaches, we designed a comprehensive single-cell transcriptomic study. Time-resolved single-cell transcriptomics of the endothelial and co-evolving mural cells gave insights into cell type development, stability, and plasticity. Transfer to a 3D hydrogel microenvironment induced neovascularization and facilitated tracing of migrating, coalescing, and tubulogenic endothelial cell states. During maturation, we monitored two pericyte subtypes evolving mural cells. Profiling cell-cell interactions between pericytes and endothelial cells revealed angiogenic signals during tubulogenesis. In silico discovered ligands were tested for their capability to attract endothelial cells. Our data, analyses, and results provide an in vitro roadmap to guide vascularization in future tissue engineering.
Topics: Humans; Endothelial Cells; Induced Pluripotent Stem Cells; Neovascularization, Physiologic; Coculture Techniques; Neovascularization, Pathologic; Pericytes
PubMed: 37714147
DOI: 10.1016/j.stemcr.2023.08.008 -
Stem Cell Reviews and Reports Oct 2023In the last two decades, considerable progress has been made in the derivation of mammalian germ cells from pluripotent stem cells such as Embryonic Stem Cells (ESCs)...
In the last two decades, considerable progress has been made in the derivation of mammalian germ cells from pluripotent stem cells such as Embryonic Stem Cells (ESCs) and induced Pluripotent Stem Cells (iPSCs). The pluripotent stem cells are generally first induced into pre-gastrulating endoderm/mesoderm-like status and then specified into putative primordial germ cells (PGCs) termed PGC-like cells (PGCLCs) which possess the potential to generate oocytes and sperms. Adipose-derived mesenchymal stromal cells (ASCs) are multipotent cells, having the capacity to differentiate into cell types such as adipocytes, osteocytes and chondrocytes. Since no information is available about the capability of female human ASCs (hASCs) to generate PGCLCs, we compared protocols to produce such cells from hASCs themselves or from hASC-derived iPSCs. The results showed that, providing pre-induction into a peri-gastrulating endoderm/mesoderm-like status, hASCs can generate PGCLCs. This process, however, shows a lower efficiency than when hASC-derived iPSCs are used as starting cells. Although hASCs possess multipotency and express mesodermal genes, direct induction into PGCLCs resulted less efficient.
Topics: Animals; Humans; Female; Germ Cells; Pluripotent Stem Cells; Embryonic Stem Cells; Induced Pluripotent Stem Cells; Mesenchymal Stem Cells; Mammals
PubMed: 37338786
DOI: 10.1007/s12015-023-10561-x -
BioRxiv : the Preprint Server For... Oct 2023During heart development, a well-characterized network of transcription factors initiates cardiac gene expression and defines the precise timing and location of cardiac...
During heart development, a well-characterized network of transcription factors initiates cardiac gene expression and defines the precise timing and location of cardiac progenitor specification. However, our understanding of the post-initiation transcriptional events that regulate cardiac gene expression is still incomplete. The PAF1C component Rtf1 is a transcription regulatory protein that modulates pausing and elongation of RNA Pol II, as well as cotranscriptional histone modifications. Here we report that Rtf1 is essential for cardiogenesis in fish and mammals, and that in the absence of Rtf1 activity, cardiac progenitors arrest in an immature state. We found that Rtf1's Plus3 domain, which confers interaction with the transcriptional pausing and elongation regulator Spt5, was necessary for cardiac progenitor formation. ChIP-seq analysis further revealed changes in the occupancy of RNA Pol II around the transcription start site (TSS) of cardiac genes in morphants reflecting a reduction in transcriptional pausing. Intriguingly, inhibition of pause release in morphants and mutants restored the formation of cardiac cells and improved Pol II occupancy at the TSS of key cardiac genes. Our findings highlight the crucial role that transcriptional pausing plays in promoting normal gene expression levels in a cardiac developmental context.
PubMed: 37873297
DOI: 10.1101/2023.10.13.562296 -
International Journal of Molecular... Aug 2023The tumor microenvironment (TME) is complex and involves many different cell types that seemingly work together in helping cancer cells evade immune monitoring and... (Review)
Review
The tumor microenvironment (TME) is complex and involves many different cell types that seemingly work together in helping cancer cells evade immune monitoring and survive therapy. The advent of single-cell sequencing has greatly increased our knowledge of the cell types present in the tumor microenvironment and their role in the developing cancer. This, coupled with clinical data showing that cancer development and the response to therapy may be influenced by drugs that indirectly influence the tumor environment, highlights the need to better understand how the cells present in the TME work together. This review looks at the different cell types (cancer cells, cancer stem cells, endothelial cells, pericytes, adipose cells, cancer-associated fibroblasts, and neuronal cells) in the bladder tumor microenvironment. Their impact on immune activation and on shaping the microenvironment are discussed as well as the effects of hypertensive drugs and anesthetics on bladder cancer.
Topics: Humans; Urinary Bladder; Tumor Microenvironment; Endothelial Cells; Neoplasms; Urinary Bladder Neoplasms; Pericytes
PubMed: 37569686
DOI: 10.3390/ijms241512311 -
Advanced Science (Weinheim,... Oct 2023Hematogenous metastasis is the main approach for colorectal cancer liver metastasis (CRCLM). However, as the gatekeepers in the tumor vessels, the role of TPCs in...
Hematogenous metastasis is the main approach for colorectal cancer liver metastasis (CRCLM). However, as the gatekeepers in the tumor vessels, the role of TPCs in hematogenous metastasis remains largely unknown, which may be attributed to the lack of specific biomarkers for TPC isolation. Here, microdissection combined with a pericyte medium-based approach is developed to obtain TPCs from CRC patients. Proteomic analysis reveals that TRP channel-associated factor 2 (TCAF2), a partner protein of the transient receptor potential cation channel subfamily M member 8 (TRPM8), is overexpressed in TPCs from patients with CRCLM. TCAF2 in TPCs is correlated with liver metastasis, short overall survival, and disease-free survival in CRC patients. Gain- and loss-of-function experiments validate that TCAF2 in TPCs promotes tumor cell motility, epithelial-mesenchymal transition (EMT), and CRCLM, which is attenuated in pericyte-conditional Tcaf2-knockout mice. Mechanistically, TCAF2 inhibits the expression and activity of TRPM8, leading to Wnt5a secretion in TPCs, which facilitates EMT via the activation of the STAT3 signaling pathway in tumor cells. Menthol, a TRPM8 agonist, significantly suppresses Wnt5a secretion in TPCs and CRCLM. This study reveals the previously unidentified pro-metastatic effects of TPCs from the perspective of cold-sensory receptors, providing a promising diagnostic biomarker and therapeutic target for CRCLM.
Topics: Mice; Animals; Humans; Pericytes; Proteomics; Thermosensing; Liver Neoplasms; Colorectal Neoplasms; TRPM Cation Channels; Membrane Proteins
PubMed: 37635201
DOI: 10.1002/advs.202302717 -
Journal of Atherosclerosis and... Sep 2023There are still many patients suffering from ischemic stroke and related disabilities worldwide. To develop a treatment that promotes functional recovery after acute... (Review)
Review
There are still many patients suffering from ischemic stroke and related disabilities worldwide. To develop a treatment that promotes functional recovery after acute ischemic stroke, we need to elucidate endogenous tissue repair mechanisms. The concept of a neurovascular unit (NVU) indicates the importance of a complex orchestration of cell-cell interactions and their microenvironment in the physiology and pathophysiology of various central nervous system diseases, particularly ischemic stroke. In this concept, microvascular pericytes play a crucial role in regulating the blood-brain barrier integrity, cerebral blood flow (CBF), and vascular stability. Recent evidence suggests that pericytes are also involved in the tissue repair leading to functional recovery following acute ischemic stroke through the interaction with other cell types constituting the NVU; pericytes may organize CBF recovery, macrophage-mediated clearance of myelin debris, intrainfarct fibrosis, and periinfarct astrogliosis and remyelination. In this review, we will discuss the physiological and pathophysiological functions of pericytes, their involvement in the molecular mechanisms underlying tissue repair and functional recovery after ischemic stroke, and a therapeutic strategy to promote endogenous regeneration.
Topics: Humans; Pericytes; Ischemic Stroke; Stroke; Blood-Brain Barrier; Macrophages
PubMed: 37394570
DOI: 10.5551/jat.RV22007 -
Stem Cell Research & Therapy Oct 2023Understanding mechanisms and manifestations of cardiovascular risk factors, including diabetes, on vascular cells such as endothelial cells, pericytes, and vascular... (Review)
Review
Understanding mechanisms and manifestations of cardiovascular risk factors, including diabetes, on vascular cells such as endothelial cells, pericytes, and vascular smooth muscle cells, remains elusive partly due to the lack of appropriate disease models. Therefore, here we explore different aspects for the development of advanced 3D in vitro disease models that recapitulate human blood vessel complications using patient-derived induced pluripotent stem cells, which retain the epigenetic, transcriptomic, and metabolic memory of their patient-of-origin. In this review, we highlight the superiority of 3D blood vessel organoids over conventional 2D cell culture systems for vascular research. We outline the key benefits of vascular organoids in both health and disease contexts and discuss the current challenges associated with organoid technology, providing potential solutions. Furthermore, we discuss the diverse applications of vascular organoids and emphasize the importance of incorporating all relevant cellular components in a 3D model to accurately recapitulate vascular pathophysiology. As a specific example, we present a comprehensive overview of diabetic vasculopathy, demonstrating how the interplay of different vascular cell types is critical for the successful modelling of complex disease processes in vitro. Finally, we propose a strategy for creating an organ-specific diabetic vasculopathy model, serving as a valuable template for modelling other types of vascular complications in cardiovascular diseases by incorporating disease-specific stressors and organotypic modifications.
Topics: Humans; Endothelial Cells; Organoids; Induced Pluripotent Stem Cells; Pericytes; Diabetes Mellitus
PubMed: 37817281
DOI: 10.1186/s13287-023-03521-2 -
Molecular Biology of the Cell May 2024The regulation of the cytoskeleton by multiple signaling pathways, sometimes in parallel, is a common principle of morphogenesis. A classic example of regulation by...
The regulation of the cytoskeleton by multiple signaling pathways, sometimes in parallel, is a common principle of morphogenesis. A classic example of regulation by parallel pathways is gastrulation, where the inputs from the Folded gastrulation (Fog)/Concertina (Cta) and the T48 pathways induce apical constriction and mesoderm invagination. Whether there are distinct roles for these separate pathways in regulating the complex spatial and temporal patterns of cytoskeletal activity that accompany early embryo development is still poorly understood. We investigated the roles of the Fog/Cta and T48 pathways and found that, by themselves, the Cta and T48 pathways both promote timely mesoderm invagination and apical myosin II accumulation, with Cta being required for timely cell shape change ahead of mitotic cell division. We also identified distinct functions of T48 and Cta in regulating cellularization and the uniformity of the apical myosin II network, respectively. Our results demonstrate that both redundant and distinct functions for the Fog/Cta and T48 pathways exist.
Topics: Animals; Drosophila; Gastrulation; Drosophila Proteins; Morphogenesis; Mesoderm; Myosin Type II; Drosophila melanogaster
PubMed: 38536475
DOI: 10.1091/mbc.E24-02-0050 -
Indian Journal of Ophthalmology May 2024The caruncle is a unique anatomical site in the human body, comprising various structures derived from the surface ectoderm and mesoderm. Caruncular lesions can range...
PURPOSE
The caruncle is a unique anatomical site in the human body, comprising various structures derived from the surface ectoderm and mesoderm. Caruncular lesions can range from benign to malignant and present challenges in accurate diagnosis and timely management due to their hidden nature and proximity to the lacrimal sac. This study aims to provide a comprehensive description of caruncular lesions, presenting the first Indian case series on this topic.
METHODS
Ethical approval was obtained, and data collection was conducted at a tertiary care center in India. A retrospective analysis was performed on 44 patients with caruncular lesions treated between 2013 and 2020. Detailed patient histories, clinical examinations, slit lamp imaging, and excision biopsies were conducted. Histopathological examination of the specimens was carried out.
RESULTS
The study included 42 cases of caruncular lesions, with a mean age of 31.09 years. The majority of cases were male (54.54%). Benign lesions accounted for 84.09% of the cases, while premalignant and malignant lesions accounted for 11.36% and 4.54%, respectively. Papilloma and nevus were the most common lesions, with 11 cases each. All caruncular lesions were successfully and completely excised without complications. Histopathological examination confirmed the accuracy of the diagnoses, with an 84.09% concordance rate between clinical assessment and pathological diagnosis.
CONCLUSION
This case series reveals a predominance of benign lesions among individuals in their early thirties. The successful excision of all lesions with a high concordance rate between clinical assessment and histopathological diagnosis underscores the importance of timely and accurate management.
PubMed: 38770617
DOI: 10.4103/IJO.IJO_2088_23