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Scientific Reports Mar 2017The combination of microfluidics with engineered three-dimensional (3D) matrices can bring new insights into the fate regulation of stem cells and their...
The combination of microfluidics with engineered three-dimensional (3D) matrices can bring new insights into the fate regulation of stem cells and their self-organization into organoids. Although there has been progress in 3D stem cell culturing, most existing in vitro methodologies do not allow for mimicking of the spatiotemporal heterogeneity of stimuli that drive morphogenetic processes in vivo. To address this, we present a perfusion-free microchip concept for the in vitro 3D perturbation of stem cell fate. Stem cells are encapsulated in a hydrogel compartment that is flanked by open reservoirs for the diffusion-driven generation of biomolecule gradients. Juxtaposing additional compartments bearing supportive cells enables investigating the influence of long range cell-cell communication. We explore the utility of the microchips in manipulating early fate choices and self-organizing characteristics of 3D-cultured mouse embryonic stem cells (mESCs) under neural differentiation conditions and exposure to gradients of leukemia inhibitory factor (LIF). mESCs respond to LIF gradients in a spatially dependent manner. At higher LIF concentrations, multicellular colonies maintain pluripotency in contrast, at lower concentrations, mESCs develop into apicobasally polarized epithelial cysts. This versatile system can help to systematically explore the role of multifactorial microenvironments in promoting self-patterning of various stem cell types.
Topics: Animals; Cell Lineage; Cells, Cultured; Cellular Microenvironment; Diffusion; Extracellular Matrix; Leukemia Inhibitory Factor; Mice; Microfluidics; Microtechnology; Mouse Embryonic Stem Cells; Pluripotent Stem Cells; Solubility
PubMed: 28303935
DOI: 10.1038/srep44711 -
Stem Cells Translational Medicine Sep 2021StemCellQC is a video bioinformatics software tool for the quantitative analysis of human pluripotent stem cell (hPSC) colonies. Our objective was to use StemCellQC to...
StemCellQC is a video bioinformatics software tool for the quantitative analysis of human pluripotent stem cell (hPSC) colonies. Our objective was to use StemCellQC to evaluate and compare various experimental culture conditions, cell lines, and treatments and to demonstrate its applicability to PSC problems. Seven key features were identified that provided useful information on PSC morphology, dynamic behavior, and viability. Colony attachment was better on laminin-521 than on Matrigel and Geltrex. Growth rates were similar on each matrix when data were normalized. The brightness/area ratio feature showed greater cell death in colonies grown on Matrigel and Geltrex than on laminin-521 further contributing to an overall greater yield of cells on laminin-521. Four different PSC culture media performed similarly; however, one medium produced batch-to-batch variation in colony morphology and dynamic features. Two embryonic and one induced pluripotent stem cell line showed significant differences in morphology, growth rates, motility, and death rates. Cells from the same vial that became phenotypically different in culture showed measurable differences in morphology, brightness, and motility. Likewise, differentiating and undifferentiated colonies varied in growth rate, intensity, and motility. Three pluripotent cell lines treated with a low concentration of cinnamaldehyde, a chemical used in consumer products, showed adverse effects and differed in their sensitivity to treatment. Our data demonstrate various applications of StemCellQC which could be used in basic and translational research, toxicological and drug testing, and clinical facilities engaged in stem cell therapy.
Topics: Cell Culture Techniques; Cell Differentiation; Computational Biology; Humans; Induced Pluripotent Stem Cells; Pluripotent Stem Cells
PubMed: 34089307
DOI: 10.1002/sctm.15-0352 -
Stem Cells Translational Medicine Oct 2020Heart diseases (HDs) are the leading cause of morbidity and mortality worldwide. Despite remarkable clinical progress made, current therapies cannot restore the lost...
Heart diseases (HDs) are the leading cause of morbidity and mortality worldwide. Despite remarkable clinical progress made, current therapies cannot restore the lost myocardium, and the correlation of genotype to phenotype of many HDs is poorly modeled. In the past two decades, with the rapid developments of human pluripotent stem cell (hPSC) biology and technology that allow the efficient preparation of cardiomyocytes from individual patients, tremendous efforts have been made for using hPSC-derived cardiomyocytes in preclinical and clinical cardiac therapy as well as in dissection of HD mechanisms to develop new methods for disease prediction and treatment. However, their applications have been hampered by several obstacles. Here, we discuss recent advances, remaining challenges, and the potential solutions to advance this field.
Topics: Cell Culture Techniques; Cell Differentiation; Embryonic Stem Cells; Heart Diseases; Humans; Myocytes, Cardiac; Pluripotent Stem Cells; Regeneration
PubMed: 32725800
DOI: 10.1002/sctm.19-0340 -
Cells Oct 2020Embryoid bodies (EBs) resemble self-organizing aggregates of pluripotent stem cells that recapitulate some aspects of early embryogenesis. Within few days, the cells... (Review)
Review
Embryoid bodies (EBs) resemble self-organizing aggregates of pluripotent stem cells that recapitulate some aspects of early embryogenesis. Within few days, the cells undergo a transition from rather homogeneous epithelial-like pluripotent stem cell colonies into a three-dimensional organization of various cell types with multifaceted cell-cell interactions and lumen formation-a process associated with repetitive epithelial-mesenchymal transitions. In the last few years, culture methods have further evolved to better control EB size, growth, cellular composition, and organization-e.g., by the addition of morphogens or different extracellular matrix molecules. There is a growing perception that the mechanical properties, cell mechanics, and cell signaling during EB development are also influenced by physical cues to better guide lineage specification; substrate elasticity and topography are relevant, as well as shear stress and mechanical strain. Epithelial structures outside and inside EBs support the integrity of the cell aggregates and counteract mechanical stress. Furthermore, hydrogels can be used to better control the organization and lineage-specific differentiation of EBs. In this review, we summarize how EB formation is accompanied by a variety of biomechanical parameters that need to be considered for the directed and reproducible self-organization of early cell fate decisions.
Topics: Biomechanical Phenomena; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; Embryoid Bodies; Embryonic Development; Humans; Pluripotent Stem Cells
PubMed: 33050550
DOI: 10.3390/cells9102270 -
Stem Cell Research Oct 2020The Spanish National Stem Cell Bank (Banco Nacional de Líneas Celulares, BNLC) was established in 2006 thanks to a change in the legislative framework in Spain. The Law...
The Spanish National Stem Cell Bank (Banco Nacional de Líneas Celulares, BNLC) was established in 2006 thanks to a change in the legislative framework in Spain. The Law 14/2006 updated the previous Assisted Reproduction Techniques Law (Law 45/2003) allowing the use of the surplus frozen embryos following IVF for research. The BNLC has a network structure with 3 nodes: the Regenerative Medicine Program (IDIBELL), the Principe Felipe Research Center (CIPF) in Valencia and the Andalusian Public Health System Biobank (SSPA Biobank) in Granada. The aim of the BNLC is to guarantee throughout the national territory the availability of human stem cell lines for biomedical research. At present time, there are 40 human embryonic stem cell lines (hESC) and 171 human induced pluripotent stem cell lines (hiPSC) registered in the BNLC. These lines are fully characterized and available in the context of research projects approved by the Technical Committee of the BNLC.
Topics: Cell Differentiation; Cell Line; Embryonic Stem Cells; Government Regulation; Humans; Induced Pluripotent Stem Cells; Pluripotent Stem Cells; Spain; Tissue Banks
PubMed: 32905997
DOI: 10.1016/j.scr.2020.101956 -
Stem Cell Reviews and Reports Feb 2015Recent advances on human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) have brought us closer... (Review)
Review
Recent advances on human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) have brought us closer to the realization of their clinical potential. Nonetheless, tissue engineering and regenerative medicine applications will require the generation of hPSC products well beyond the laboratory scale. This also mandates the production of hPSC therapeutics in fully-defined, xeno-free systems and in a reproducible manner. Toward this goal, we summarize current developments in defined media free of animal-derived components for hPSC culture. Bioinspired and synthetic extracellular matrices for the attachment, growth and differentiation of hPSCs are also reviewed. Given that most progress in xeno-free medium and substrate development has been demonstrated in two-dimensional rather than three dimensional culture systems, translation from the former to the latter poses unique difficulties. These challenges are discussed in the context of cultivation platforms of hPSCs as aggregates, on microcarriers or after encapsulation in biocompatible scaffolds.
Topics: Animals; Cell Culture Techniques; Cell Proliferation; Cellular Microenvironment; Embryonic Stem Cells; Extracellular Matrix; Humans; Induced Pluripotent Stem Cells; Models, Biological; Pluripotent Stem Cells
PubMed: 25077810
DOI: 10.1007/s12015-014-9544-x -
Future Medicinal Chemistry Jun 2019Most neurodegenerative diseases are characterized by a complex and mostly still unresolved pathology. This fact, together with the lack of reliable disease models, has... (Review)
Review
Most neurodegenerative diseases are characterized by a complex and mostly still unresolved pathology. This fact, together with the lack of reliable disease models, has precluded the development of effective therapies counteracting the disease progression. The advent of human pluripotent stem cells has revolutionized the field allowing the generation of disease-relevant neural cell types that can be used for disease modeling, drug screening and, possibly, cell transplantation purposes. In this Review, we discuss the applications of human pluripotent stem cells, the development of efficient protocols for the derivation of the different neural cells and their applicability for robust disease modeling and drug screening platforms for most common neurodegenerative conditions.
Topics: Animals; CRISPR-Cas Systems; Drug Evaluation, Preclinical; Gene Editing; Humans; Neurodegenerative Diseases; Neurogenesis; Neurons; Pluripotent Stem Cells
PubMed: 31161803
DOI: 10.4155/fmc-2018-0520 -
Trends in Cell Biology May 2017The advent of human pluripotent stem cell (hPSC) biology has opened unprecedented opportunities for the use of tissue engineering to generate human cardiac tissue for in... (Review)
Review
The advent of human pluripotent stem cell (hPSC) biology has opened unprecedented opportunities for the use of tissue engineering to generate human cardiac tissue for in vitro study. Engineering cardiac constructs that recapitulate human development and disease requires faithful recreation of the cardiac niche in vitro. Here we discuss recent progress in translating the in vivo cardiac microenvironment into PSC models of the human heart. We review three key physiologic features required to recreate the cardiac niche and facilitate normal cardiac differentiation and maturation: the biochemical, biophysical, and bioelectrical signaling cues. Finally, we discuss key barriers that must be overcome to fulfill the promise of stem cell biology in preclinical applications and ultimately in clinical practice.
Topics: Cellular Microenvironment; Disease; Humans; Models, Biological; Physiological Phenomena; Pluripotent Stem Cells
PubMed: 28007424
DOI: 10.1016/j.tcb.2016.11.010 -
Trends in Molecular Medicine Sep 2019Pluripotent stem cells (PSCs) represent an attractive cell source for treating muscular dystrophies (MDs) since they easily allow for the generation of large numbers of... (Review)
Review
Pluripotent stem cells (PSCs) represent an attractive cell source for treating muscular dystrophies (MDs) since they easily allow for the generation of large numbers of highly regenerative myogenic progenitors. Using reprogramming technology, patient-specific PSCs have been derived for several types of MDs, and genome editing has allowed correction of mutations, opening the opportunity for their therapeutic application in an autologous transplantation setting. However, there has been limited progress on preclinical studies that validate the therapeutic potential of these gene corrected PSC-derived myogenic progenitors. In this review, we highlight the major research advances, challenges, and future prospects towards the development of PSC-based therapeutics for MDs.
Topics: Animals; Biomarkers; Cell Differentiation; Cell- and Tissue-Based Therapy; Disease Models, Animal; Gene Editing; Genetic Therapy; Humans; Induced Pluripotent Stem Cells; Muscular Dystrophies; Myoblasts, Cardiac; Pluripotent Stem Cells; Regeneration; Stem Cell Transplantation; Transgenes
PubMed: 31473142
DOI: 10.1016/j.molmed.2019.07.004 -
Stem Cell Research May 2021The Korea National Stem Cell Bank has been banking pluripotent stem cell (PSC) lines since 2012. Quality-controlled and ethically sourced cell lines were developed for...
The Korea National Stem Cell Bank has been banking pluripotent stem cell (PSC) lines since 2012. Quality-controlled and ethically sourced cell lines were developed for distribution. Currently (as of 2020), among the 69 deposited lines, 4 research-grade human embryonic stem cell (hESC) lines and 19 induced pluripotent stem cell (iPSC) lines have been distributed. Good manufacturing practices (GMP)-compliant homozygous iPSC lines for regenerative medicine with homozygous HLA haplotypes that cover 51% of the Korean population have been deposited as well. To ensure the quality of the cell lines, we performed eighteen different quality tests on the identity, sterility, consistency, stability and safety of the cell lines. Regarding genetic stability, we are collecting SNPchip, WES, Methyl-seq, and RNA-seq data, which are open to the public.
Topics: Cell Line; Embryonic Stem Cells; Humans; Induced Pluripotent Stem Cells; Pluripotent Stem Cells; Republic of Korea
PubMed: 33714852
DOI: 10.1016/j.scr.2021.102270