Neural commitment of human pluripotent stem cells under defined conditions recapitulates neural development and generates patient-specific neural cells

2015 ◽  
Vol 10 (10) ◽  
pp. 1578-1588 ◽  
Author(s):  
Tiago G. Fernandes ◽  
Sofia T. Duarte ◽  
Mehrnaz Ghazvini ◽  
Cláudia Gaspar ◽  
Diana C. Santos ◽  
...  
Keyword(s):  
Stem Cells ◽  
Neural Development ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Patient Specific ◽  
Neural Cells

scholarly journals Development of a modular automated system for maintenance and differentiation of adherent human pluripotent stem cells

2016 ◽  
Author(s):  
Duncan E. Crombie ◽  
Maciej Daniszewski ◽  
Helena H. Liang ◽  
Tejal Kulkarni ◽  
Fan Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
Human Pluripotent Stem Cells ◽  
Automated System ◽  
Patient Specific ◽  
High Throughput Analysis ◽  
Large Scale Culture ◽  
Induced Pluripotent ◽  
Selection Of

AbstractPatient-specific induced pluripotent stem cells (iPSCs) have tremendous potential for development of regenerative medicine, disease modelling and drug discovery. However, the processes of reprogramming, maintenance and differentiation are labour intensive and subject to inter-technician variability. To address these issues, we established and optimised protocols to allow for the automated maintenance of reprogrammed somatic cells into iPSCs to enable the large-scale culture and passaging of human pluripotent stem cells (PSCs) using a customized TECAN Freedom EVO. Generation of iPSCs was performed offline by nucleofection followed by selection of TRA-1-60 positive cells using a Miltenyi MultiMACS24 Separator. Pluripotency markers were assessed to confirm pluripotency of the generated iPSCs. Passaging was performed using an enzyme-free dissociation method. Proof of concept of differentiation was obtained by differentiating human PSCs into cells of the retinal lineage. Key advantages of this automated approach are the ability to increase sample size, reduce variability during reprogramming or differentiation, and enable medium to high-throughput analysis of human PSCs and derivatives. These techniques will become increasingly important with the emergence of clinical trials using stem cells.

Human Pluripotent Stem Cells in Neurodegenerative Diseases: Potentials, Advances and Limitations

2020 ◽  
Vol 15 (2) ◽  
pp. 102-110 ◽  
Author(s):  
Tannaz Akbari Kolagar ◽  
Maryam Farzaneh ◽  
Negin Nikkar ◽  
Seyed Esmaeil Khoshnam
Keyword(s):  
Stem Cells ◽  
Neurodegenerative Diseases ◽  
Pluripotent Stem Cells ◽  
Neurodegenerative Disorders ◽  
Disease Modeling ◽  
Human Pluripotent Stem Cells ◽  
Neural Regeneration ◽  
Patient Specific ◽  
Human Escs ◽  
Human Ipscs

Neurodegenerative diseases are progressive and uncontrolled gradual loss of motor neurons function or death of neuron cells in the central nervous system (CNS) and the mechanisms underlying their progressive nature remain elusive. There is urgent need to investigate therapeutic strategies and novel treatments for neural regeneration in disorders like Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and multiple sclerosis (MS). Currently, the development and identification of pluripotent stem cells enabling the acquisition of a large number of neural cells in order to improve cell recovery after neurodegenerative disorders. Pluripotent stem cells which consist of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are characterized by their ability to indefinitely self-renew and the capacity to differentiate into different types of cells. The first human ESC lines were established from donated human embryos; while, because of a limited supply of donor embryos, human ESCs derivation remains ethically and politically controversial. Hence, hiPSCs-based therapies have been shown as an effective replacement for human ESCs without embryo destruction. Compared to the invasive methods for derivation of human ESCs, human iPSCs has opened possible to reprogram patient-specific cells by defined factors and with minimally invasive procedures. Human pluripotent stem cells are a good source for cell-based research, cell replacement therapies and disease modeling. To date, hundreds of human ESC and human iPSC lines have been generated with the aim of treating various neurodegenerative diseases. In this review, we have highlighted the recent potentials, advances, and limitations of human pluripotent stem cells for the treatment of neurodegenerative disorders.

scholarly journals Dorsal-ventral patterned neural cyst from human pluripotent stem cells in a neurogenic niche

2019 ◽  
Vol 5 (12) ◽  
pp. eaax5933 ◽  
Author(s):  
Y. Zheng ◽  
X. Xue ◽  
A. M. Resto-Irizarry ◽  
Z. Li ◽  
Y. Shao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Neural Development ◽  
Pluripotent Stem Cells ◽  
System Development ◽  
Nervous System Development ◽  
Human Pluripotent Stem Cells ◽  
Human Embryos ◽  
Neurogenic Niche ◽  
Developmental Divergence ◽  
Organizing Principles

Despite its importance in central nervous system development, development of the human neural tube (NT) remains poorly understood, given the challenges of studying human embryos, and the developmental divergence between humans and animal models. We report a human NT development model, in which NT-like tissues, neuroepithelial (NE) cysts, are generated in a bioengineered neurogenic environment through self-organization of human pluripotent stem cells (hPSCs). NE cysts correspond to the neural plate in the dorsal ectoderm and have a default dorsal identity. Dorsal-ventral (DV) patterning of NE cysts is achieved using retinoic acid and/or sonic hedgehog and features sequential emergence of the ventral floor plate, P3, and pMN domains in discrete, adjacent regions and a dorsal territory progressively restricted to the opposite dorsal pole. This hPSC-based, DV patterned NE cyst system will be useful for understanding the self-organizing principles that guide NT patterning and for investigations of neural development and neural disease.

scholarly journals A Step-by-Step Refined Strategy for Highly Efficient Generation of Neural Progenitors and Motor Neurons from Human Pluripotent Stem Cells

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3087
Author(s):  
Jie Ren ◽  
Chaoyi Li ◽  
Mengfei Zhang ◽  
Huakun Wang ◽  
Yali Xie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Motor Neurons ◽  
Neural Development ◽  
Pluripotent Stem Cells ◽  
Disease Modeling ◽  
Human Pluripotent Stem Cells ◽  
Highly Efficient ◽  
Efficient Generation ◽  
Human Neurons

Limited access to human neurons, especially motor neurons (MNs), was a major challenge for studying neurobiology and neurological diseases. Human pluripotent stem cells (hPSCs) could be induced as neural progenitor cells (NPCs) and further multiple neural subtypes, which provide excellent cellular sources for studying neural development, cell therapy, disease modeling and drug screening. It is thus important to establish robust and highly efficient methods of neural differentiation. Enormous efforts have been dedicated to dissecting key signalings during neural commitment and accordingly establishing reliable differentiation protocols. In this study, we refined a step-by-step strategy for rapid differentiation of hPSCs towards NPCs within merely 18 days, combining the adherent and neurosphere-floating methods, as well as highly efficient generation (~90%) of MNs from NPCs by introducing refined sets of transcription factors for around 21 days. This strategy made use of, and compared, retinoic acid (RA) induction and dual-SMAD pathway inhibition, respectively, for neural induction. Both methods could give rise to highly efficient and complete generation of preservable NPCs, but with different regional identities. Given that the generated NPCs can be differentiated into the majority of excitatory and inhibitory neurons, but hardly MNs, we thus further differentiate NPCs towards MNs by overexpressing refined sets of transcription factors, especially by adding human SOX11, whilst improving a series of differentiation conditions to yield mature MNs for good modeling of motor neuron diseases. We thus refined a detailed step-by-step strategy for inducing hPSCs towards long-term preservable NPCs, and further specified MNs based on the NPC platform.

scholarly journals Development of a Modular Automated System for Maintenance and Differentiation of Adherent Human Pluripotent Stem Cells

2017 ◽  
Vol 22 (8) ◽  
pp. 1016-1025 ◽  
Author(s):  
Duncan E. Crombie ◽  
Maciej Daniszewski ◽  
Helena H. Liang ◽  
Tejal Kulkarni ◽  
Fan Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
Disease Modeling ◽  
Human Pluripotent Stem Cells ◽  
Automated System ◽  
Patient Specific ◽  
High Throughput Analysis ◽  
Large Scale Culture ◽  
Induced Pluripotent

Patient-specific induced pluripotent stem cells (iPSCs) have tremendous potential for development of regenerative medicine, disease modeling, and drug discovery. However, the processes of reprogramming, maintenance, and differentiation are labor intensive and subject to intertechnician variability. To address these issues, we established and optimized protocols to allow for the automated maintenance of reprogrammed somatic cells into iPSCs to enable the large-scale culture and passaging of human pluripotent stem cells (PSCs) using a customized TECAN Freedom EVO. Generation of iPSCs was performed offline by nucleofection followed by selection of TRA-1-60–positive cells using a Miltenyi MultiMACS24 Separator. Pluripotency markers were assessed to confirm pluripotency of the generated iPSCs. Passaging was performed using an enzyme-free dissociation method. Proof of concept of differentiation was obtained by differentiating human PSCs into cells of the retinal lineage. Key advantages of this automated approach are the ability to increase sample size, reduce variability during reprogramming or differentiation, and enable medium- to high-throughput analysis of human PSCs and derivatives. These techniques will become increasingly important with the emergence of clinical trials using stem cells.

scholarly journals Building on a Solid Foundation: Adding Relevance and Reproducibility to Neurological Modeling Using Human Pluripotent Stem Cells

2021 ◽  
Vol 15 ◽  
Author(s):  
Erin Knock ◽  
Lisa M. Julian
Keyword(s):  
Stem Cells ◽  
Animal Models ◽  
Human Brain ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Model Systems ◽  
Model Organisms ◽  
Slice Culture ◽  
Neural Cells ◽  
Cell Culture Techniques

The brain is our most complex and least understood organ. Animal models have long been the most versatile tools available to dissect brain form and function; however, the human brain is highly distinct from that of standard model organisms. In addition to existing models, access to human brain cells and tissues is essential to reach new frontiers in our understanding of the human brain and how to intervene therapeutically in the face of disease or injury. In this review, we discuss current and developing culture models of human neural tissue, outlining advantages over animal models and key challenges that remain to be overcome. Our principal focus is on advances in engineering neural cells and tissue constructs from human pluripotent stem cells (PSCs), though primary human cell and slice culture are also discussed. By highlighting studies that combine animal models and human neural cell culture techniques, we endeavor to demonstrate that clever use of these orthogonal model systems produces more reproducible, physiological, and clinically relevant data than either approach alone. We provide examples across a range of topics in neuroscience research including brain development, injury, and cancer, neurodegenerative diseases, and psychiatric conditions. Finally, as testing of PSC-derived neurons for cell replacement therapy progresses, we touch on the advancements that are needed to make this a clinical mainstay.

scholarly journals Modeling Rett Syndrome with Human Pluripotent Stem Cells: Mechanistic Outcomes and Future Clinical Perspectives

2021 ◽  
Vol 22 (7) ◽  
pp. 3751
Author(s):  
Ana Rita Gomes ◽  
Tiago G. Fernandes ◽  
Joaquim M.S. Cabral ◽  
Maria Margarida Diogo
Keyword(s):  
Stem Cells ◽  
Rett Syndrome ◽  
Pluripotent Stem Cells ◽  
Neurodevelopmental Disorder ◽  
Human Pluripotent Stem Cells ◽  
Screening Tests ◽  
Patient Specific ◽  
Promising Alternative ◽  
The Impact

Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the gene encoding the methyl-CpG-binding protein 2 (MeCP2). Among many different roles, MeCP2 has a high phenotypic impact during the different stages of brain development. Thus, it is essential to intensively investigate the function of MeCP2, and its regulated targets, to better understand the mechanisms of the disease and inspire the development of possible therapeutic strategies. Several animal models have greatly contributed to these studies, but more recently human pluripotent stem cells (hPSCs) have been providing a promising alternative for the study of RTT. The rapid evolution in the field of hPSC culture allowed first the development of 2D-based neuronal differentiation protocols, and more recently the generation of 3D human brain organoid models, a more complex approach that better recapitulates human neurodevelopment in vitro. Modeling RTT using these culture platforms, either with patient-specific human induced pluripotent stem cells (hiPSCs) or genetically-modified hPSCs, has certainly contributed to a better understanding of the onset of RTT and the disease phenotype, ultimately allowing the development of high throughput drugs screening tests for potential clinical translation. In this review, we first provide a brief summary of the main neurological features of RTT and the impact of MeCP2 mutations in the neuropathophysiology of this disease. Then, we provide a thorough revision of the more recent advances and future prospects of RTT modeling with human neural cells derived from hPSCs, obtained using both 2D and organoids culture systems, and its contribution for the current and future clinical trials for RTT.

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