Stem Cell

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    Protocols in Current Issue
    Generation of T cells from Human and Nonhuman Primate Pluripotent Stem Cells
    [Abstract] Pluripotent stem cells (PSCs) have the potential to provide homogeneous cell populations of T cells that can be grown at a clinical scale and genetically engineered to meet specific clinical needs. OP9-DLL4, a stromal line ectopically expressing the Notch ligand Delta-like 4 (DLL4) is used to support differentiation of PSCs to T-lymphocytes. This ...
    In vivo Mouse Mammary Gland Formation
    Authors:  Heng Sun, Xin Zhang, Un In Chan, Sek Man Su, Sen Guo, Xiaoling Xu and Chuxia Deng, date: 07/05/2020, view: 538, Q&A: 0
    [Abstract] For years, the mammary gland serves as a perfect example to study the self-renew and differentiation of adult stem cells, and the regulatory mechanisms of these processes as well. To assess the function of given genes and/or other factors on stemness of mammary cells, several In vitro assays were developed, such as mammospheres formation ...
    In vitro Self-organized Mouse Small Intestinal Epithelial Monolayer Protocol
    Authors:  Gizem Altay, Eduard Batlle, Vanesa Fernández-Majada and Elena Martinez, date: 02/05/2020, view: 2469, Q&A: 0
    [Abstract] Developing protocols to obtain intestinal epithelial monolayers that recapitulate in vivo physiology to overcome the limitations of the organoids’ closed geometry has become of great interest during the last few years. Most of the developed culture models showed physiological-relevant cell composition but did not prove self-renewing ...
    Retina Injury and Retina Tissue Preparation to Study Regeneration in Zebrafish
    Authors:  Poonam Sharma and Rajesh Ramachandran, date: 12/20/2019, view: 1501, Q&A: 0
    [Abstract] Unlike mammals, primitive vertebrates have immense capability to regenerate almost all of their organs including the central nervous system. Among primitive organisms, zebrafish have been extensively used as a model system for regeneration studies. The retina is a part of the central nervous system and mammals lack the potential to repair any ...
    In vitro Differentiation of Human iPSC-derived Retinal Pigment Epithelium Cells (iPSC-RPE)
    Authors:  Agnieszka D’Antonio-Chronowska, Matteo D’Antonio and Kelly A. Frazer, date: 12/20/2019, view: 2265, Q&A: 0
    [Abstract] Induced Pluripotent Stem Cells (iPSCs) serve as an excellent model system for studying the molecular underpinnings of tissue development. Human iPSC-derived retinal pigment epithelium (iPSC-RPE) cells have fetal-like molecular profiles. Hence, biobanks like iPSCORE, which contain iPSCs generated from hundreds of individuals, are an invaluable ...
    Mapping the Mechanome–A Protocol for Simultaneous Live Imaging and Quantitative Analysis of Cell Mechanoadaptation and Ingression
    [Abstract] Mechanomics, the mechanics equivalent of genomics, is a burgeoning field studying mechanical modulation of stem cell behavior and lineage commitment. Analogous to mechanical testing of a living material as it adapts and evolves, mapping of the mechanome necessitates the development of new protocols to assess changes in structure and function in ...
    Co-culture of Human Stem Cell Derived Neurons and Oligodendrocyte Progenitor Cells
    Authors:  Stephanie Dooves, Aishwarya G. Nadadhur, Lisa Gasparotto and Vivi M. Heine, date: 09/05/2019, view: 2398, Q&A: 0
    [Abstract] Crosstalk between neurons and oligodendrocytes is important for proper brain functioning. Multiple co-culture methods have been developed to study oligodendrocyte maturation, myelination or the effect of oligodendrocytes on neurons. However, most of these methods contain cells derived from animal models. In the current protocol, we co-culture ...
    Axon-seq for in Depth Analysis of the RNA Content of Neuronal Processes
    Authors:  Jik Nijssen, Julio Aguila and Eva Hedlund, date: 07/20/2019, view: 4066, Q&A: 0
    [Abstract] Neuronal processes have an RNA composition that is distinct from the cell body. Therefore, to fully understand neuronal biology in health and disease we need to study both somas, dendrites and axons. Here we describe a detailed protocol of a newly refined method, Axon-seq, for RNA sequencing of axons (and dendrites) grown in isolation using single ...
    CRISPR/Cas9 + AAV-mediated Intra-embryonic Gene Knocking in Mice
    [Abstract] Intra-embryo genome editing by CRISPR/Cas9 has enabled rapid generation of gene knockout animals. However, large fragment knock-in directly into embryos’ genome is still difficult, especially without microinjection of donor DNA. Viral vectors are good transporters of knock-in donor DNA for cell lines, but seemed unsuitable for pre-implantation ...
    Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) into an Effective Model of Forebrain Neural Progenitor Cells and Mature Neurons
    [Abstract] Induced Pluripotent Stem Cells (iPSCs) are pluripotent stem cells that can be generated from somatic cells, and provide a way to model the development of neural tissues in vitro. One particularly interesting application of iPSCs is the development of neurons analogous to those found in the human forebrain. Forebrain neurons play a central ...



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