Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6040180

Title:In vitro generation of differentiated neurons from cultures of mammalian multipotential CNS stem cells
Inventors:Johe, Karl K.; Potomac, MD, USA
Summary:This invention introduces methods of propagation and the long-term in vitro culturing of central nervous system stem cells, such that the cells retain their differentiation capacity and exhibit selective differentiation into neurons, astrocytes and oligodendrocytes. Described herein are region-specific, terminally differentiated, mature neurons derived from cultures of mammalian multipotential CNS stem cells and an in vitro procedure by which the differentiated neurons may be generated. Further disclosed are methods by which neurons, astrocytes and oligodendrocytes may be generated from such an adhesion culture of mammalian multipotential CNS stem cells in a serum-free medium supplemented with a mitogen, the withdrawal of which renders such differentiation. Methods of harvesting the stem cells by trypsinization are disclosed, as are conditions under which the stem cells should be optimally plated at a density of between 100,000 to 250,000 cells per square centimeter. Therapeutic applications are included in gene and cell therapy and in the screening of various drugs and growth factors.
Abstract:The present invention reveals in vitro cultures of region-specific, terminally differentiated, mature neurons derived from cultures of mammalian multipotential CNS stem cells and an in vitro procedure by which the differentiated neurons may be generated. The procedure involves the culturing of multipotential CNS stem cells from a specific region in a chemically defined serum-free culture medium containing a growth factor; replacing the medium with growth factor-free medium; harvesting the stem cells by trypsinization; plating the stem cells at a density of between 100,000 to 250,000 cells per square centimeter; and culturing the stem cells in a glutamic acid-free chemically defined serum-free culture medium.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6040180
Application Number:US1997000919580
Date Filed:07/05/1997
Date Published:21/03/2000
Assignee:NeuralStem Biopharmaceuticals, Ltd., College Park, MD, USA


 
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