scholarly journals Niche-guided tissue patterning by chemomechanical flow lithography

2021 ◽  
Author(s):  
Peter L. H. Newman ◽  
Pierre Osteil ◽  
Tim A. Anderson ◽  
Jane Q. J. Sun ◽  
Daryan Kempe ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Cell Types ◽  
Germ Layer ◽  
Young’S Moduli ◽  
Tissue Patterning ◽  
Organ Systems ◽  
Cell Source ◽  
And Function ◽  
Tissue Models

Pluripotent-stem-cell-derived tissue-models have been established with increasingly physiological shape, size and function1–3. However, the histogenic and morphogenetic processes present in these models proceed stochastically. This reflects an absence of technologies able to produce complex supportive cell niches that can reproducibly guide tissue patterning and generate well-defined tissue structures. To address this, we have developed chemomechanical flow lithography (CMFL), a printing technology that delivers orthogonally programmable chemical and mechanical properties to microstructured niches that drive the differentiation of selective cell types and spatial emplacement of these cells in a micropattern. We print microstructured niches conjugated with peptides, proteins and macromolecular morphogens across a range of Young’s moduli. Using such niches, we generated tissue patterned constructs from a single cell source with regionalised cell differentiation, including a bone-fat osteoid from stromal mesenchyme, and a patterned assembly of germ-layer tissues derived from pluripotent stem cells. Thus, CMFL is a valuable tool for generating orthogonally and chemomechanically defined niches able to guide spatially defined tissue patterns. This approach enables studies to better understand how extrinsic niche factors regulate histogenic and morphogenetic processes, towards engineering complex structured tissue and organ systems with higher-level emergent function.

scholarly journals Present and future challenges of induced pluripotent stem cells

2015 ◽  
Vol 370 (1680) ◽  
pp. 20140367 ◽  
Author(s):  
Mari Ohnuki ◽  
Kazutoshi Takahashi
Keyword(s):  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Disease Modelling ◽  
Future Perspectives ◽  
Differentiated Cells ◽  
Human Ipscs ◽  
Future Challenges ◽  
Induced Pluripotent

Growing old is our destiny. However, the mature differentiated cells making up our body can be rejuvenated to an embryo-like fate called pluripotency which is an ability to differentiate into all cell types by enforced expression of defined transcription factors. The discovery of this induced pluripotent stem cell (iPSC) technology has opened up unprecedented opportunities in regenerative medicine, disease modelling and drug discovery. In this review, we introduce the applications and future perspectives of human iPSCs and we also show how iPSC technology has evolved along the way.

scholarly journals An orthogonal differentiation platform for genomically programming stem cells, organoids, and bioprinted tissues

2020 ◽  
Author(s):  
Mark A. Skylar-Scott ◽  
Jeremy Y. Huang ◽  
Aric Lu ◽  
Alex H.M. Ng ◽  
Tomoya Duenki ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Pluripotent Stem Cells ◽  
Cell Types ◽  
Embryoid Bodies ◽  
One Pot ◽  
Neural Tissues ◽  
Induced Pluripotent ◽  
And Function ◽  
Matrix Free

AbstractSimultaneous differentiation of human induced pluripotent stem cells (hiPSCs) into divergent cell types offers a pathway to achieving tailorable cellular complexity, patterned architecture, and function in engineered human organoids and tissues. Recent transcription factor (TF) overexpression protocols typically produce only one cell type of interest rather than the multitude of cell types and structural organization found in native human tissues. Here, we report an orthogonal differentiation platform for genomically programming stem cells, organoids and bioprinted tissues with controlled composition and organization. To demonstrate this platform, we orthogonally differentiated endothelial cells and neurons from hiPSCs in a one-pot system containing neural stem cell-specifying media. By aggregating inducible-TF and wildtype hiPSCs into pooled and multicore-shell embryoid bodies, we produced vascularized and patterned cortical organoids within days. Using multimaterial 3D bioprinting, we patterned 3D neural tissues from densely cellular, matrix-free stem cell inks that were orthogonally differentiated on demand into distinct layered regions composed of neural stem cells, endothelium, and neurons, respectively. Given the high proliferative capacity and patient-specificity of hiPSCs, our platform provides a facile route for programming cells and multicellular tissues for drug screening and therapeutic applications.

scholarly journals Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors

2017 ◽  
Vol 2017 ◽  
pp. 1-15 ◽  
Author(s):  
Casey L. Roberts ◽  
Silvia S. Chen ◽  
Angela C. Murchison ◽  
Rebecca A. Ogle ◽  
Michael P. Francis ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cell Types ◽  
Embryoid Bodies ◽  
Population Doubling ◽  
Specific Cell ◽  
Germ Layer ◽  
Induced Pluripotent ◽  
Endodermal Cells

While induced pluripotent stem cells (iPSCs) hold great clinical promise, one hurdle that remains is the existence of a parental germ-layer memory in reprogrammed cells leading to preferential differentiation fates. While it is problematic for generating cells vastly different from the reprogrammed cells’ origins, it could be advantageous for the reliable generation of germ-layer specific cell types for future therapeutic use. Here we use human osteoblast-derived iPSCs (hOB-iPSCs) to generate induced osteoprogenitors (iOPs). Osteoblasts were successfully reprogrammed and demonstrated by endogenous upregulation of Oct4, Sox2, Nanog, TRA-1-81, TRA-16-1, SSEA3, and confirmatory hPSC Scorecard Algorithmic Assessment. The hOB-iPSCs formed embryoid bodies with cells of ectoderm and mesoderm but have low capacity to form endodermal cells. Differentiation into osteoprogenitors occurred within only 2–6 days, with a population doubling rate of less than 24 hrs; however, hOB-iPSC derived osteoprogenitors were only able to form osteogenic and chondrogenic cells but not adipogenic cells. Consistent with this, hOB-iOPs were found to have higher methylation of PPARγbut similar levels of methylation on the RUNX2 promoter. These data demonstrate that iPSCs can be generated from human osteoblasts, but variant methylation patterns affect their differentiation capacities. Therefore, epigenetic memory can be exploited for efficient generation of clinically relevant quantities of osteoprogenitor cells.

scholarly journals Human Pluripotent Stem Cells-Based Therapies for Neurodegenerative Diseases: Current Status and Challenges

Cells ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 2517
Author(s):  
Elizabeth Ford ◽  
Jodie Pearlman ◽  
Travis Ruan ◽  
John Manion ◽  
Matthew Waller ◽  
...  
Keyword(s):  
Stem Cells ◽  
Nervous System ◽  
Stem Cell ◽  
Neurodegenerative Diseases ◽  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Cell Types ◽  
Cell Therapies ◽  
Wide Range ◽  
Cord Injury

Neurodegenerative diseases are characterized by irreversible cell damage, loss of neuronal cells and limited regeneration potential of the adult nervous system. Pluripotent stem cells are capable of differentiating into the multitude of cell types that compose the central and peripheral nervous systems and so have become the major focus of cell replacement therapies for the treatment of neurological disorders. Human embryonic stem cell (hESC) and human induced pluripotent stem cell (hiPSC)-derived cells have both been extensively studied as cell therapies in a wide range of neurodegenerative disease models in rodents and non-human primates, including Parkinson’s disease, stroke, epilepsy, spinal cord injury, Alzheimer’s disease, multiple sclerosis and pain. In this review, we discuss the latest progress made with stem cell therapies targeting these pathologies. We also evaluate the challenges in clinical application of human pluripotent stem cell (hPSC)-based therapies including risk of oncogenesis and tumor formation, immune rejection and difficulty in regeneration of the heterogeneous cell types composing the central nervous system.

scholarly journals Remdesivir but not famotidine inhibits SARS-CoV-2 replication in human pluripotent stem cell-derived intestinal organoids

2020 ◽  
Author(s):  
Jana Krüger ◽  
Rüdiger Groß ◽  
Carina Conzelmann ◽  
Janis A. Müller ◽  
Lennart Koepke ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Gastrointestinal Symptoms ◽  
Cell Types ◽  
Human Pluripotent Stem Cell ◽  
Intestinal Organoids

Gastrointestinal symptoms in COVID-19 are associated with prolonged symptoms and increased severity. We employed human intestinal organoids derived from pluripotent stem cells (PSC-HIOs) to analyze SARS-CoV-2 pathogenesis and to validate efficacy of specific drugs in the gut. Certain, but not all cell types in PSC-HIOs express SARS-CoV-2 entry factors ACE2 and TMPRSS2, rendering them susceptible to SARS-CoV-2 infection. Remdesivir, a promising drug to treat COVID-19, effectively suppressed SARS-CoV-2 infection of PSC-HIOs. In contrast, the histamine-2-blocker famotidine showed no effect. Thus, PSC-HIOs provide an interesting platform to study SARS-CoV-2 infection and to identify or validate drugs.

scholarly journals Stepwise in vitro induction of human somitic mesoderm and its derivatives

2020 ◽  
Author(s):  
Yoshihiro Yamanaka ◽  
Maya Uemura ◽  
Cantas Alev
Keyword(s):  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Developmental Process ◽  
Cell Types ◽  
Model Organisms ◽  
Human Segmentation ◽  
Induced Pluripotent ◽  
Somitic Mesoderm ◽  
In Vitro Induction

Abstract Our understanding of human somitogenesis is limited and largely based on insights gained from model organisms. Pluripotent stem cell-based in vitro approaches aiming to recapitulate distinct aspects of this core developmental process have recently been reported, including our recent paper on the in vitro recapitulation of the human segmentation clock1. Here we describe in detail our stepwise induction protocol of presomitic mesoderm (PSM), somitic mesoderm (SM), and its two major derivatives, sclerotome (SCL) and dermomyotome (DM) from human induced pluripotent stem cells (iPSCs). We further briefly address the subsequent molecular and functional analysis of these in vitro induced human mesodermal lineages and cell-types.

scholarly journals Effect of Human Donor Cell Source on Differentiation and Function of Cardiac Induced Pluripotent Stem Cells

2014 ◽  
Vol 64 (5) ◽  
pp. 436-448 ◽  
Author(s):  
Veronica Sanchez-Freire ◽  
Andrew S. Lee ◽  
Shijun Hu ◽  
Oscar J. Abilez ◽  
Ping Liang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Donor Cell ◽  
Human Donor ◽  
Cell Source ◽  
Induced Pluripotent ◽  
And Function

scholarly journals Stepwise in vitro induction of human somitic mesoderm and its derivatives

2020 ◽  
Author(s):  
Cantas Alev ◽  
Yoshihiro Yamanaka ◽  
Maya Uemura
Keyword(s):  
Pluripotent Stem Cells ◽  
Pluripotent Stem Cell ◽  
Developmental Process ◽  
Cell Types ◽  
Model Organisms ◽  
Human Segmentation ◽  
Induced Pluripotent ◽  
Somitic Mesoderm ◽  
In Vitro Induction

Abstract Our understanding of human somitogenesis is limited and largely based on insights gained from model organisms. Pluripotent stem cell-based in vitro approaches aiming to recapitulate distinct aspects of this core developmental process have recently been reported, including our recent paper on the in vitro recapitulation of the human segmentation clock 1 . Here we describe in detail our stepwise induction protocol of presomitic mesoderm (PSM), somitic mesoderm (SM), and its two major derivatives, sclerotome (SCL) and dermomyotome (DM) from human induced pluripotent stem cells (iPSCs). We further briefly address the subsequent molecular and functional analysis of these in vitro induced human mesodermal lineages and cell-types.

scholarly journals Single cell determination of cardiac microtissue structure and function using light sheet microscopy

2020 ◽  
Author(s):  
Diwakar Turaga ◽  
Oriane B. Matthys ◽  
Tracy A. Hookway ◽  
David A. Joy ◽  
Meredith Calvert ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Single Cell ◽  
Cardiac Tissue ◽  
Imaging Techniques ◽  
Light Sheet ◽  
Cell Types ◽  
The Individual ◽  
And Function ◽  
Tissue Models ◽  
Light Sheet Fluorescence Microscopy

AbstractNative cardiac tissue is comprised of heterogeneous cell populations that work cooperatively for proper tissue function; thus, engineered tissue models have moved toward incorporating multiple cardiac cell types in an effort to recapitulate native multicellular composition and organization. Cardiac tissue models comprised of stem cell-derived cardiomyocytes require inclusion of non-myocytes to promote stable tissue formation, yet the specific contributions of the supporting non-myocyte population on the parenchymal cardiomyocytes and cardiac microtissues have yet to be fully dissected. This gap can be partly attributed to limitations in technologies able to accurately study the individual cellular structure and function that comprise intact 3D tissues. The ability to interrogate the cell-cell interactions in 3D tissue constructs has been restricted by conventional optical imaging techniques that fail to adequately penetrate multicellular microtissues with sufficient spatial resolution. Light sheet fluorescence microscopy overcomes these constraints to enable single cell-resolution structural and functional imaging of intact cardiac microtissues. Multicellular spatial distribution analysis of heterotypic cardiac cell populations revealed that cardiomyocytes and cardiac fibroblasts were randomly distributed throughout 3D microtissues. Furthermore, calcium imaging of live cardiac microtissues enabled single-cell detection of cardiomyocyte calcium activity, which showed that functional heterogeneity correlated with spatial location within the tissues. This study demonstrates that light sheet fluorescence microscopy can be utilized to determine single-cell spatial and functional interactions of multiple cell types within intact 3D engineered microtissues, thereby facilitating the determination of structure-function relationships at both tissue-level and single-cell resolution.Impact StatementThe ability to achieve single-cell resolution by advanced 3D light imaging techniques enables exquisite new investigation of multicellular analyses in native and engineered tissues. In this study, light sheet fluorescence microscopy was used to define structure-function relationships of distinct cell types in engineered cardiac microtissues by determining heterotypic cell distributions and interactions throughout the tissues as well as by assessing regional differences in calcium handing functional properties at the individual cardiomyocyte level.

Role of the 807 C/T Polymorphism of the α2 Gene in Platelet GP Ia Collagen Receptor Expression and Function

1999 ◽  
Vol 81 (06) ◽  
pp. 951-956 ◽  
Author(s):  
J. Corral ◽  
R. González-Conejero ◽  
J. Rivera ◽  
F. Ortuño ◽  
P. Aparicio ◽  
...  
Keyword(s):  
Venous Thrombosis ◽  
Cell Types ◽  
Receptor Expression ◽  
Case Control Studies ◽  
Platelet Surface ◽  
Vitro Analysis ◽  
And Function ◽  
In Vitro Analysis

SummaryThe variability of the platelet GP Ia/IIa density has been associated with the 807 C/T polymorphism (Phe 224) of the GP Ia gene in American Caucasian population. We have investigated the genotype and allelic frequencies of this polymorphism in Spanish Caucasians. The T allele was found in 35% of the 284 blood donors analyzed. We confirmed in 159 healthy subjects a significant association between the 807 C/T polymorphism and the platelet GP Ia density. The T allele correlated with high number of GP Ia molecules on platelet surface. In addition, we observed a similar association of this polymorphism with the expression of this protein in other blood cell types. The platelet responsiveness to collagen was determined by “in vitro” analysis of the platelet activation and aggregation response. We found no significant differences in these functional platelet parameters according to the 807 C/T genotype. Finally, results from 3 case/control studies involving 302 consecutive patients (101 with coronary heart disease, 104 with cerebrovascular disease and 97 with deep venous thrombosis) determined that the 807 C/T polymorphism of the GP Ia gene does not represent a risk factor for arterial or venous thrombosis.

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