The Experts below are selected from a list of 19629 Experts worldwide ranked by ideXlab platform
Donald E Ingber - One of the best experts on this subject based on the ideXlab platform.
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Directed Differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a glomerulus chip
Nature Protocols, 2018Co-Authors: Samira Musah, Diogo M Camacho, Nikolaos Dimitrakakis, Donald E Ingber, George M ChurchAbstract:Protocols have been established to direct the Differentiation of human induced pluripotent stem (iPS) cells into nephron progenitor cells and organoids containing many types of kidney cells, but it has been difficult to direct the Differentiation of iPS cells to form specific types of mature human kidney cells with high yield. Here, we describe a detailed protocol for the Directed Differentiation of human iPS cells into mature, post-mitotic kidney glomerular podocytes with high (>90%) efficiency within 26 d and under chemically defined conditions, without genetic manipulations or subpopulation selection. We also describe how these iPS cell–derived podocytes may be induced to form within a microfluidic organ-on-a-chip (Organ Chip) culture device to build a human kidney Glomerulus Chip that mimics the structure and function of the kidney glomerular capillary wall in vitro within 35 d (starting with undifferentiated iPS cells). The podocyte Differentiation protocol requires skills for culturing iPS cells, and the development of a Glomerulus Chip requires some experience with building and operating microfluidic cell culture systems. This method could be useful for applications in nephrotoxicity screening, therapeutic development, and regenerative medicine, as well as mechanistic study of kidney development and disease. Ingber and colleagues describe how to generate kidney glomerular podocytes from human pluripotent stem cells and how to engineer a human kidney Glomerulus Chip.
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Directed Differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a glomerulus chip
Nature Protocols, 2018Co-Authors: Samira Musah, Diogo M Camacho, Nikolaos Dimitrakakis, Donald E Ingber, George M ChurchAbstract:Protocols have been established to direct the Differentiation of human induced pluripotent stem (iPS) cells into nephron progenitor cells and organoids containing many types of kidney cells, but it has been difficult to direct the Differentiation of iPS cells to form specific types of mature human kidney cells with high yield. Here, we describe a detailed protocol for the Directed Differentiation of human iPS cells into mature, post-mitotic kidney glomerular podocytes with high (>90%) efficiency within 26 d and under chemically defined conditions, without genetic manipulations or subpopulation selection. We also describe how these iPS cell-derived podocytes may be induced to form within a microfluidic organ-on-a-chip (Organ Chip) culture device to build a human kidney Glomerulus Chip that mimics the structure and function of the kidney glomerular capillary wall in vitro within 35 d (starting with undifferentiated iPS cells). The podocyte Differentiation protocol requires skills for culturing iPS cells, and the development of a Glomerulus Chip requires some experience with building and operating microfluidic cell culture systems. This method could be useful for applications in nephrotoxicity screening, therapeutic development, and regenerative medicine, as well as mechanistic study of kidney development and disease.
Egor Y Plotnikov - One of the best experts on this subject based on the ideXlab platform.
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human induced pluripotent stem cells derived from fetal neural stem cells successfully undergo Directed Differentiation into cartilage
Stem Cells and Development, 2011Co-Authors: S P Medvedev, E V Grigoreva, Alexander I Shevchenko, A A Malakhova, Elena V Dementyeva, Alexander A Shilov, Evgeny Pokushalov, Alla M Zaidman, M A Aleksandrova, Egor Y PlotnikovAbstract:Induced pluripotent stem (iPS) cells can be derived from a wide range of somatic cells via overexpression of a set of specific genes. With respect to their properties, iPS cells closely resemble embryonic stem cells. Because of their main property, pluripotency, iPS cells have excellent prospects for use in substitutive cell therapy; however, the methods of Directed Differentiation of iPS cells have not been yet sufficiently elaborated. In this work, we derived human iPS cells from fetal neural stem (FNS) cells by transfection with a polycistronic plasmid vector carrying the mouse Oct4, Sox2, Klf4, and c-Myc genes or a plasmid expressing the human OCT4 gene. We have shown that human FNS cells can be effectively reprogrammed despite a low transfection level (10%–15%) and that the use of 2-propylvaleric (valproic) acid and BIX-01294 increases the yield of iPS cell clones to ∼7-fold. Further, transient expression of OCT4 alone is sufficient for reprogramming. The iPS cells obtained express all the major mark...
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human induced pluripotent stem cells derived from fetal neural stem cells successfully undergo Directed Differentiation into cartilage
Stem Cells and Development, 2011Co-Authors: S P Medvedev, E V Grigoreva, Alexander I Shevchenko, A A Malakhova, Elena V Dementyeva, Alexander A Shilov, Evgeny Pokushalov, Alla M Zaidman, M A Aleksandrova, Egor Y PlotnikovAbstract:Induced pluripotent stem (iPS) cells can be derived from a wide range of somatic cells via overexpression of a set of specific genes. With respect to their properties, iPS cells closely resemble embryonic stem cells. Because of their main property, pluripotency, iPS cells have excellent prospects for use in substitutive cell therapy; however, the methods of Directed Differentiation of iPS cells have not been yet sufficiently elaborated. In this work, we derived human iPS cells from fetal neural stem (FNS) cells by transfection with a polycistronic plasmid vector carrying the mouse Oct4, Sox2, Klf4, and c-Myc genes or a plasmid expressing the human OCT4 gene. We have shown that human FNS cells can be effectively reprogrammed despite a low transfection level (10%-15%) and that the use of 2-propylvaleric (valproic) acid and BIX-01294 increases the yield of iPS cell clones to ∼7-fold. Further, transient expression of OCT4 alone is sufficient for reprogramming. The iPS cells obtained express all the major markers of embryonic stem cells and are able to differentiate in vitro into ectodermal, mesodermal, and endodermal derivatives. In addition, we have found that the human iPS cells derived from FNS cells can be successfully subjected to in vitro Directed chondrogenic Differentiation to form functional cartilaginous tissue.
S P Medvedev - One of the best experts on this subject based on the ideXlab platform.
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human induced pluripotent stem cells derived from fetal neural stem cells successfully undergo Directed Differentiation into cartilage
Stem Cells and Development, 2011Co-Authors: S P Medvedev, E V Grigoreva, Alexander I Shevchenko, A A Malakhova, Elena V Dementyeva, Alexander A Shilov, Evgeny Pokushalov, Alla M Zaidman, M A Aleksandrova, Egor Y PlotnikovAbstract:Induced pluripotent stem (iPS) cells can be derived from a wide range of somatic cells via overexpression of a set of specific genes. With respect to their properties, iPS cells closely resemble embryonic stem cells. Because of their main property, pluripotency, iPS cells have excellent prospects for use in substitutive cell therapy; however, the methods of Directed Differentiation of iPS cells have not been yet sufficiently elaborated. In this work, we derived human iPS cells from fetal neural stem (FNS) cells by transfection with a polycistronic plasmid vector carrying the mouse Oct4, Sox2, Klf4, and c-Myc genes or a plasmid expressing the human OCT4 gene. We have shown that human FNS cells can be effectively reprogrammed despite a low transfection level (10%–15%) and that the use of 2-propylvaleric (valproic) acid and BIX-01294 increases the yield of iPS cell clones to ∼7-fold. Further, transient expression of OCT4 alone is sufficient for reprogramming. The iPS cells obtained express all the major mark...
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human induced pluripotent stem cells derived from fetal neural stem cells successfully undergo Directed Differentiation into cartilage
Stem Cells and Development, 2011Co-Authors: S P Medvedev, E V Grigoreva, Alexander I Shevchenko, A A Malakhova, Elena V Dementyeva, Alexander A Shilov, Evgeny Pokushalov, Alla M Zaidman, M A Aleksandrova, Egor Y PlotnikovAbstract:Induced pluripotent stem (iPS) cells can be derived from a wide range of somatic cells via overexpression of a set of specific genes. With respect to their properties, iPS cells closely resemble embryonic stem cells. Because of their main property, pluripotency, iPS cells have excellent prospects for use in substitutive cell therapy; however, the methods of Directed Differentiation of iPS cells have not been yet sufficiently elaborated. In this work, we derived human iPS cells from fetal neural stem (FNS) cells by transfection with a polycistronic plasmid vector carrying the mouse Oct4, Sox2, Klf4, and c-Myc genes or a plasmid expressing the human OCT4 gene. We have shown that human FNS cells can be effectively reprogrammed despite a low transfection level (10%-15%) and that the use of 2-propylvaleric (valproic) acid and BIX-01294 increases the yield of iPS cell clones to ∼7-fold. Further, transient expression of OCT4 alone is sufficient for reprogramming. The iPS cells obtained express all the major markers of embryonic stem cells and are able to differentiate in vitro into ectodermal, mesodermal, and endodermal derivatives. In addition, we have found that the human iPS cells derived from FNS cells can be successfully subjected to in vitro Directed chondrogenic Differentiation to form functional cartilaginous tissue.
Samira Musah - One of the best experts on this subject based on the ideXlab platform.
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Directed Differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a glomerulus chip
Nature Protocols, 2018Co-Authors: Samira Musah, Diogo M Camacho, Nikolaos Dimitrakakis, Donald E Ingber, George M ChurchAbstract:Protocols have been established to direct the Differentiation of human induced pluripotent stem (iPS) cells into nephron progenitor cells and organoids containing many types of kidney cells, but it has been difficult to direct the Differentiation of iPS cells to form specific types of mature human kidney cells with high yield. Here, we describe a detailed protocol for the Directed Differentiation of human iPS cells into mature, post-mitotic kidney glomerular podocytes with high (>90%) efficiency within 26 d and under chemically defined conditions, without genetic manipulations or subpopulation selection. We also describe how these iPS cell–derived podocytes may be induced to form within a microfluidic organ-on-a-chip (Organ Chip) culture device to build a human kidney Glomerulus Chip that mimics the structure and function of the kidney glomerular capillary wall in vitro within 35 d (starting with undifferentiated iPS cells). The podocyte Differentiation protocol requires skills for culturing iPS cells, and the development of a Glomerulus Chip requires some experience with building and operating microfluidic cell culture systems. This method could be useful for applications in nephrotoxicity screening, therapeutic development, and regenerative medicine, as well as mechanistic study of kidney development and disease. Ingber and colleagues describe how to generate kidney glomerular podocytes from human pluripotent stem cells and how to engineer a human kidney Glomerulus Chip.
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Directed Differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a glomerulus chip
Nature Protocols, 2018Co-Authors: Samira Musah, Diogo M Camacho, Nikolaos Dimitrakakis, Donald E Ingber, George M ChurchAbstract:Protocols have been established to direct the Differentiation of human induced pluripotent stem (iPS) cells into nephron progenitor cells and organoids containing many types of kidney cells, but it has been difficult to direct the Differentiation of iPS cells to form specific types of mature human kidney cells with high yield. Here, we describe a detailed protocol for the Directed Differentiation of human iPS cells into mature, post-mitotic kidney glomerular podocytes with high (>90%) efficiency within 26 d and under chemically defined conditions, without genetic manipulations or subpopulation selection. We also describe how these iPS cell-derived podocytes may be induced to form within a microfluidic organ-on-a-chip (Organ Chip) culture device to build a human kidney Glomerulus Chip that mimics the structure and function of the kidney glomerular capillary wall in vitro within 35 d (starting with undifferentiated iPS cells). The podocyte Differentiation protocol requires skills for culturing iPS cells, and the development of a Glomerulus Chip requires some experience with building and operating microfluidic cell culture systems. This method could be useful for applications in nephrotoxicity screening, therapeutic development, and regenerative medicine, as well as mechanistic study of kidney development and disease.
Suchun Zhang - One of the best experts on this subject based on the ideXlab platform.
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Directed Differentiation of forebrain GABA interneurons from human pluripotent stem cells
Nature Protocols, 2013Co-Authors: Conall Sauvey, Ewa D Zarnowska, Suchun ZhangAbstract:Forebrain γ-aminobutyric acid (GABA) interneurons have crucial roles in high-order brain function via modulating network activities and plasticity, and they are implicated in many psychiatric disorders. Availability of enriched functional human forebrain GABA interneurons, especially those from people affected by GABA interneuron deficit disease, will be instrumental to the investigation of disease pathogenesis and development of therapeutics. We describe a protocol for Directed Differentiation of forebrain GABA interneurons from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in a chemically defined system. In this protocol, human PSCs are first induced to primitive neuroepithelial cells over 10 d, and then patterned to NKX2.1-expressing medial ganglionic eminence progenitors by simple treatment with sonic hedgehog or its agonist purmorphamine over the next 2 weeks. These progenitors generate a nearly pure population of forebrain GABA interneurons by the sixth week. This simple and efficient protocol does not require transgenic modification or cell sorting, and it has been replicated with multiple human ESC and iPSC lines.
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Directed Differentiation of functional astroglial subtypes from human pluripotent stem cells
Nature Protocols, 2011Co-Authors: Robert Krencik, Suchun ZhangAbstract:Directed Differentiation of functional astroglial subtypes from human pluripotent stem cells
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Directed Differentiation of functional astroglial subtypes from human pluripotent stem cells
Nature Protocols, 2011Co-Authors: Robert Krencik, Suchun ZhangAbstract:Regionally and functionally diverse types of astrocytes exist throughout the central nervous system and participate in nearly every aspect of normal and abnormal neural function. Therefore, human astrocyte subtypes are useful tools for understanding brain function, modulating disease processes and promoting neural regeneration. Here we describe a protocol for Directed Differentiation and maintenance of functional astroglia from human pluripotent stem cells in a chemically defined system. Human stem cells are first differentiated into neuroepithelial cells with or without exogenous patterning molecules (days 0–21). Regular dissociation of the neuroepithelial clusters in suspension, and in the presence of mitogens, permits generation of astroglial subtypes over a long-term expansion (days 21–90). Finally, the astroglial progenitors are either amplified for an extended time or differentiated into functional astrocytes on removal of mitogens and the addition of ciliary neurotrophic factor (days >90). This method generates robust populations of functionally diversified astrocytes with high efficiency.
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Directed Differentiation of neural stem cells and subtype specific neurons from hescs
Methods of Molecular Biology, 2010Co-Authors: Suchun ZhangAbstract:We describe a chemically defined protocol for efficient Differentiation of human embryonic stem cells (hESCs) to neural epithelial cells and then to functional spinal motor neurons. This protocol comprises four major steps. Human ESCs are differentiated without morphogens into neuroepithelial cells that form neural tube-like rosettes in the first 2 weeks. The neuroepithelial cells are then specified to OLIG2-expressing motoneuron progenitors in the presence of retinoic acid (RA) and sonic hedgehog (SHH) in the following 2 weeks. These OLIG2 progenitors generate postmitotic, HB9 expressing motoneurons at the fifth week and mature to functional motor neurons thereafter. The protein factor SHH can be replaced by a small molecule purmorphamine in the entire process, which may facilitate potential clinical applications. This protocol has been shown equally effective in generating motor neurons from human induced pluropotent stem (iPS) cells.
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Directed Differentiation of dopaminergic neuronal subtypes from human embryonic stem cells
Stem Cells, 2005Co-Authors: Yiping Yan, Ewa D Zarnowska, James A Thomson, Dali Yang, Brian Werbel, Chuck Valliere, Robert A Pearce, Suchun ZhangAbstract:How dopamine (DA) neuronal subtypes are specified remains unknown. In this study we show a robust generation of functional DA neurons from human embryonic stem cells (hESCs) through a specific sequence of application of fibroblast growth factor 8 (FGF8) and sonic hedgehog (SHH). Treatment of hESC-derived Sox1+ neuroepithelial cells with FGF8 and SHH resulted in production of tyrosine hydroxylase (TH)–positive neurons that were mostly bipolar cells, coexpression with γ-aminobutyric acid, and lack of midbrain marker engrailed 1 (En1) expression. However, FGF8 treatment of precursor cells before Sox1 expression led to the generation of a similar proportion of TH+ neurons characteristic of midbrain projection DA neurons with large cell bodies and complex processes and coexpression of En1. This suggests that one mechanism of generating neuronal subtypes is temporal availability of morphogens to a specific group of precursors. The in vitro–generated DA neurons were electrophysiologically active and released DA in an activity-dependent manner. They may thus provide a renewable source of functional human DA neurons for drug screening and development of sustainable therapeutics for disorders affecting the DA system.