Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5589376

Title:Mammalian neural crest stem cells
Inventors:Anderson, David J.; Altadena, CA, USA
Stemple, Derek L.; Pasadena, CA, USA
Summary:Described herein are mammalian multipotent neural stem cells, methods and uses thereof. The invention relates to a process for the isolation and clonal propagation of mammalian neural crest stem cells which are capable of self-renewal, asymmetrical division and differentiation into neuronal and glial progenitors and their immortalized forms when grown in a culture free of feeder cells. Further disclosed are methods by which a novel separation and culturing regimen is employed in combination with bioassays for establishing the generation of neural crest stem cell derivatives. Claims of the invention include lineage restriction which is sensitive to the local environment such that neural crest stem cells cultured on a mixed substrate of poly-D-lysine and fibronectin generate PNS neurons and glia, but when cultured on fibronectin alone the stem cells generate PNS glia but not neurons. Therapeutic applications in transplantation or gene therapy for the treatment of nervous system diseases are included.
Abstract:The invention includes methods for the isolation and clonal propagation of mammalian neural crest stem cells and isolated cellular compositions comprising the same. The methods employ a novel separation and culturing regimen and bioassays for establishing the generation of neural crest stem cell derivatives. These methods result in the production of non-transformed neural crest stem cells and their progeny. The invention demonstrates, at the clonal level, the self regeneration and asymmetrical division of mammalian neural crest stem cells for the first time in feeder cell-independent cultures. Lineage restriction is demonstrated within a developing clone and is shown to be sensitive to the local environment. Neural crest stem cells cultured on a mixed substrate of poly-D-lysine and fibronectin generate PNS neurons and glia, but on fibronectin alone the stem cells generate PNS glia but not neurons. The neurogenic potential of the stem cells, while not expressed, is maintained over time on fibronectin. The invention further includes transplantation assays which allow for the identification of mammalian neural crest stem cells from various tissues. It also includes methods for transplanting mammalian neural crest stem cells and/or neural or glial progenitors into mammals.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5589376
Application Number:US1994000290228
Date Filed:15/08/1994
Date Published:31/12/1996
Assignee:California Institute of Technology, Pasadena, CA, USA


 
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