scholarly journals Human iPS-derived pre-epicardial cells direct cardiomyocyte aggregation expansion and organization in vitro

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jun Jie Tan ◽  
Jacques P. Guyette ◽  
Kenji Miki ◽  
Ling Xiao ◽  
Gurbani Kaur ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Spatial Organization ◽  
Three Dimensional ◽  
Calcium Handling ◽  
Lateral Plate Mesoderm ◽  
Lateral Plate ◽  
Epicardial Cells ◽  
Tissue Organization ◽  
Cell Layers

AbstractEpicardial formation is necessary for normal myocardial morphogenesis. Here, we show that differentiating hiPSC-derived lateral plate mesoderm with BMP4, RA and VEGF (BVR) can generate a premature form of epicardial cells (termed pre-epicardial cells, PECs) expressing WT1, TBX18, SEMA3D, and SCX within 7 days. BVR stimulation after Wnt inhibition of LPM demonstrates co-differentiation and spatial organization of PECs and cardiomyocytes (CMs) in a single 2D culture. Co-culture consolidates CMs into dense aggregates, which then form a connected beating syncytium with enhanced contractility and calcium handling; while PECs become more mature with significant upregulation of UPK1B, ITGA4, and ALDH1A2 expressions. Our study also demonstrates that PECs secrete IGF2 and stimulate CM proliferation in co-culture. Three-dimensional PEC-CM spheroid co-cultures form outer smooth muscle cell layers on cardiac micro-tissues with organized internal luminal structures. These characteristics suggest PECs could play a key role in enhancing tissue organization within engineered cardiac constructs in vitro.

Epicardial Cells of Human Adults Can Undergo an Epithelial-to-Mesenchymal Transition and Obtain Characteristics of Smooth Muscle Cells In Vitro

Stem Cells ◽  
2007 ◽  
Vol 25 (2) ◽  
pp. 271-278 ◽  
Author(s):  
John van Tuyn ◽  
Douwe E. Atsma ◽  
Elizabeth M. Winter ◽  
Ietje van der Velde-van Dijke ◽  
Daniel A. Pijnappels ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Epithelial To Mesenchymal Transition ◽  
Muscle Cells ◽  
Mesenchymal Transition ◽  
Epicardial Cells ◽  
Human Adults

scholarly journals Development of A Three-Dimensional Tissue Construct from Dental Human Ectomesenchymal Stem Cells: In Vitro and In Vivo Study

2012 ◽  
Vol 6 (1) ◽  
pp. 226-234 ◽  
Author(s):  
Daniela Guzmán-Uribe ◽  
Keila Neri Alvarado Estrada ◽  
Amaury de Jesús Pozos Guillén ◽  
Silvia Martín Pérez ◽  
Raúl Rosales Ibáñez
Keyword(s):  
Three Dimensional ◽  
Human Tooth ◽  
Animal Cells ◽  
Dental Tissue ◽  
Membrane Markers ◽  
Tissue Organization ◽  
Specific Membrane ◽  
Tissue Construct

Application of regenerative medicine technology provides treatment for patients with several clinical problems, like loss of tissue and its function. The investigation of biological tooth replacement, dental tissue engineering and cell culture, scaffolds and growth factors are considered essential. Currently, studies reported on the making of threedimensional tissue constructs focused on the use of animal cells in the early stages of embryogenesis applied to young biomodels. The purpose of this study was the development and characterization of a three-dimensional tissue construct from human dental cells. The construct was detached, cultured and characterized in mesenchymal and epithelial cells of a human tooth germ of a 12 year old patient. The cells were characterized by specific membrane markers (STRO1, CD44), making a biocomplex using Pura Matrix as a scaffold, and it was incubated for four days and transplanted into 30 adult immunosuppressed male Wistar rats. They were evaluated at 6 days, 10 days and 2 months, obtaining histological sections stained with hematoxylin and eosin. Cell cultures were positive for specific membrane markers, showing evident deviations in morphology under phase contrast microscope. Differentiation and organization were noted at 10 days, while the constructs at 2 months showed a clear difference in morphology, organization and cell type. It was possible to obtain a three-dimensional tissue construct from human dental ectomesenchymal cells achieving a degree of tissue organization that corresponds to the presence of cellular stratification and extracellular matrix.

Fabrication of in vitro three-dimensional multilayered blood vessel model using human endothelial and smooth muscle cells and high-strength PEG hydrogel

2013 ◽  
Vol 116 (2) ◽  
pp. 231-234 ◽  
Author(s):  
Sho Shinohara ◽  
Takanori Kihara ◽  
Shinji Sakai ◽  
Michiya Matsusaki ◽  
Mitsuru Akashi ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Blood Vessel ◽  
Smooth Muscle Cells ◽  
Three Dimensional ◽  
High Strength ◽  
Muscle Cells

scholarly journals Overexpression of Netrin-1 Induces Neovascularization in the Adult Mouse Brain

2008 ◽  
Vol 28 (9) ◽  
pp. 1543-1551 ◽  
Author(s):  
Yongfeng Fan ◽  
Fanxia Shen ◽  
Yongmei Chen ◽  
Qi Hao ◽  
Weizhong Liu ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Network Formation ◽  
Cell Monolayer ◽  
Muscle Cells ◽  
Tube Formation ◽  
Adult Brain ◽  
Cell Layers

Netrin-1 is a critical molecule for axonal pathfinding during embryo development, and because of its structural homology to the endothelial mitogens, it may share its effects on vascular network formation. Using an adeno-associated viral netrin-1 vector (AAV-NT-1) gene transfer, we demonstrated that netrin-1 was able to stimulate the proliferation and migration of human cerebral endothelial cells (HCECs) and human aortic smooth muscle cells (HASMCs) compared with the control ( P < 0.05), and could also promote HCEC tube formation on matrigel ( P < 0.05) in vitro. Moreover, netrin-1 hyperstimulation could promote focal neovascularization ( P < 0.05) in the adult brain in vivo. Unlike VEGF-induced microvessel increase, netrin-1-induced newly formed vessels showed an artery-like phenotype, with an intact endothelial cell monolayer surrounded by multiple cell layers, including smooth muscle cells and an astrocyte-connected outer layer. Our findings suggest that netrin-1 plays an important role in promoting blood vessel formation in the adult rodent central nervous system, and could have broad implication in cerebrovascular development and remodeling.

scholarly journals Three-dimensional migration of macrophages requires Hck for podosome organization and extracellular matrix proteolysis

Blood ◽  
2010 ◽  
Vol 115 (7) ◽  
pp. 1444-1452 ◽  
Author(s):  
Céline Cougoule ◽  
Véronique Le Cabec ◽  
Renaud Poincloux ◽  
Talal Al Saati ◽  
Jean-Louis Mège ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Spatial Organization ◽  
Ectopic Expression ◽  
Three Dimensional ◽  
Matrix Metalloproteases ◽  
3 Dimensional ◽  
Normal Expression ◽  
Migration Of Macrophages

Abstract Tissue infiltration of phagocytes exacerbates several human pathologies including chronic inflammations or cancers. However, the mechanisms involved in macrophage migration through interstitial tissues are poorly understood. We investigated the role of Hck, a Src-family kinase involved in the organization of matrix adhesion and degradation structures called podosomes. In Hck−/− mice submitted to peritonitis, we found that macrophages accumulated in interstitial tissues and barely reached the peritoneal cavity. In vitro, 3-dimensional (3D) migration and matrix degradation abilities, 2 protease-dependent properties of bone marrow–derived macrophages (BMDMs), were affected in Hck−/− BMDMs. These macrophages formed few and undersized podosome rosettes and, consequently, had reduced matrix proteolysis operating underneath despite normal expression and activity of matrix metalloproteases. Finally, in fibroblasts unable to infiltrate matrix, ectopic expression of Hck provided the gain–of–3D migration function, which correlated positively with formation of podosome rosettes. In conclusion, spatial organization of podosomes as large rosettes, proteolytic degradation of extracellular matrix, and 3D migration appeared to be functionally linked and regulated by Hck in macrophages. Hck, as the first protein combining a phagocyte-limited expression with a role in 3D migration, could be a target for new anti-inflammatory and antitumor molecules.

Behavioural properties of chick somitic mesoderm and lateral plate when explanted in vitro

Development ◽  
1980 ◽  
Vol 56 (1) ◽  
pp. 41-58
Author(s):  
Ruth Bellairs ◽  
P. A. Portch ◽  
E. J. Sanders
Keyword(s):  
Time Lapse ◽  
Lateral Plate Mesoderm ◽  
General Shape ◽  
Electron Microscopic ◽  
Lateral Plate ◽  
Plastic Substrates ◽  
Different Types ◽  
Somitic Mesoderm ◽  
Microscopic Techniques

Tissue culture, time-lapse cinematographic and electron microscopic techniques have been used to study the properties of chick mesoderm at several stages of differentiation. Lateral plate, unsegmented mesoderm (segmental plate), and newly formed somites were dissected from stage-12 embryos, whilst dermo-myotomes and sclerotomes were dissected from stage-18 embryos. Each type of mesoderm was found to exhibit a characteristic pattern of behaviour. The explants from the unsegmented mesoderm, from the newly formed somites and from the older embryos could be placed in a developmental sequence; with increasing differentiation they settled and spread on the substrate more readily, whether explanted as pieces of tissue or as individual cells, and it was concluded that this implied an increased adhesion to the substrate. Similarly, with increasing differentiation, the cells segmented at a faster rate. No significant differences could be discerned in the internal structure of the different types of cells, although differences in the general shape were apparent. The lateral plate mesoderm cells, which bear some resemblances to the unsegmented mesoderm cells in the embryo, also show some morphological resemblances to them in vitro. However, the lateral plate cells had a much greater success in attaching to glass or plastic substrates. They were also found to have the highest speed of locomotion of all the tissues studied, whereas the unsegmented had the lowest. It is concluded therefore, that although cells may look similar to one another morphologically, their behaviour may differ greatly, probably because they are already partially determined.

scholarly journals Structure of hibernating ribosomes studied by cryoelectron tomography in vitro and in situ

2010 ◽  
Vol 190 (4) ◽  
pp. 613-621 ◽  
Author(s):  
Julio O. Ortiz ◽  
Florian Brandt ◽  
Valério R.F. Matias ◽  
Lau Sennels ◽  
Juri Rappsilber ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Spatial Organization ◽  
Three Dimensional ◽  
E Coli ◽  
Escherichia Coli Cells ◽  
Three Dimensional Configuration

Ribosomes arranged in pairs (100S) have been related with nutritional stress response and are believed to represent a “hibernation state.” Several proteins have been identified that are associated with 100S ribosomes but their spatial organization has hitherto not been characterized. We have used cryoelectron tomography to reveal the three-dimensional configuration of 100S ribosomes isolated from starved Escherichia coli cells and we have described their mode of interaction. In situ studies with intact E. coli cells allowed us to demonstrate that 100S ribosomes do exist in vivo and represent an easily reversible state of quiescence; they readily vanish when the growth medium is replenished.

scholarly journals Limiting lumens: a new role for Cdc42

2008 ◽  
Vol 183 (4) ◽  
pp. 575-577 ◽  
Author(s):  
Terry Lechler
Keyword(s):  
Fluid Transport ◽  
Three Dimensional ◽  
Intestinal Lumen ◽  
Dimensional System ◽  
In Vitro System ◽  
Lumen Formation ◽  
Tissue Organization ◽  
Single Lumen ◽  
Three Dimensional System

The formation of a single lumen is a necessary step in the formation of biological tubes. Different tissues have developed diverse ways to form their lumens. In this issue, Jaffe et al. (Jaffe, A.B., N. Kaji, J. Durgan, and A. Hall. 2008. J. Cell Biol. 183:625–633) report the development of an in vitro system for studying lumen formation that is driven by fluid transport, recapitulating intestinal lumen formation. Effective ion and fluid transport requires both cell polarity and proper tissue organization. Surprisingly, polarization of cells in this three-dimensional system does not require Cdc42. Instead, Cdc42 prevents formation of multiple lumens by orienting cell divisions and directing apical membrane biogenesis.

scholarly journals Origin of congenital coronary arterio-ventricular fistulae from anomalous epicardial and myocardial development

2022 ◽  
Author(s):  
Paul Palmquist-Gomes ◽  
Adrian Ruiz-Villalba ◽  
Juan Antonio Guadix ◽  
Juan Pablo Romero ◽  
Bettina Bessieres ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Coronary Artery ◽  
Congenital Anomalies ◽  
Molecular Mechanisms ◽  
Relevant Information ◽  
Ventricular Wall ◽  
Coronary Smooth Muscle ◽  
Epicardial Cells ◽  
Developing Heart

Coronary Artery Fistulae (CAFs) are cardiac congenital anomalies consisting of an abnormal communication of a coronary artery with either a cardiac chamber or another cardiac vessel. In humans, these congenital anomalies can lead to complications such as myocardial hypertrophy, endocarditis, heart dilatation and failure. Unfortunately, despite their clinical relevance, the aetiology of CAFs remains unknown. In this work, we have used two different species (mouse and avian embryos) to experimentally model CAFs morphogenesis. Both conditional Itga4 (alpha 4 integrin) epicardial deletion in mice and cryocauterisation of chick embryonic hearts disrupted epicardial development and ventricular wall growth, two essential events in coronary embryogenesis. Additional transcriptomics and in vitro analyses were performed to better understand how arterio-ventricular connections are originated in the embryonic heart. Our results suggest myocardial discontinuities in the developing heart promote the formation of endocardial pouch-like structures resembling human CAF. The structure of these CAF-like anomalies was compared with histopathological data from a paediatric heart CAF, showing histomorphological and immunochemical similarities, including an accumulation of smooth muscle positive cells in the pouch-like structure wall. In vitro experiments showed the abnormal contact between the epicardium and the endocardium may promote the precocious differentiation of epicardial cells to smooth muscle. Our results suggest that myocardial discontinuities in the embryonic ventricular wall promote the early contact of the endocardium with epicardial-derived coronary progenitors at the cardiac surface, leading to ventricular endocardial extrusion, precocious differentiation of coronary smooth muscle cells, and the formation of pouch-like aberrant coronary-like structures in direct connection with the ventricular lumen. Our results may provide relevant information for the early diagnosis of these congenital anomalies and the molecular mechanisms that regulate their embryogenesis.

scholarly journals An organotypic in vitro model of matured blood vessels

2020 ◽  
Author(s):  
Jaehyun Lee ◽  
Esak Lee
Keyword(s):  
Smooth Muscle ◽  
Blood Vessels ◽  
Three Dimensional ◽  
Vascular Diseases ◽  
Mural Cell ◽  
Vessel Maturation ◽  
Vascular Physiology ◽  
Vitro Model ◽  
Vascular Maturation

AbstractAngiogenesis is a physiological process in which brand-new blood vessels are formed from pre-existing blood vessels. The angiogenic processes are achieved by multiple steps, including angiogenic vascular sprouting, lumen formation, mural cell (e.g., smooth muscle cells) recruitment, and vessel stabilization by the mural cell coverage of the neovessels. Especially, mural cell recruitment to and coverage of the newly formed endothelium is a fundamental process to provide fully matured, functional blood vessels. Although investigation of the mural cell interactions with endothelial cells is crucial not only for better understanding of vascular physiology, but also for treating numerous vascular diseases, there has been a lack of three-dimensional (3D) in vitro models that recapitulate spontaneous processes of the vascular maturation. In this study, we describe an organotypic in vitro model that represents multi-step, spontaneous vascular maturation processes, which includes angiogenic vessel sprouting, smooth muscle cell (SMC) recruitment, and the SMC coverage of the neovessels. Using the system, we could spatiotemporally control vessel sprouting and vessel stabilization/maturation; and revealed an optimal condition that could reconstitute SMC-covered, matured blood vessels in 3D in vitro. We may provide a new platform for future mechanism studies of vascular interactions to mural cells and vessel maturation; and for pre-clinical screening and validation of therapeutic agent candidates for treating vascular diseases.

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