Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US6667176

Title:cDNA libraries reflecting gene expression during growth and differentiation of human pluripotent stem cells
Inventors:Funk, Walter D.; Hayward, CA, USA
Carpenter, Melissa K.; Foster City, CA, USA
Gold, Joseph D.; San Francisco, CA, USA
Inokuma, Margaret S.; San Jose, CA, USA
Xu, Chunhui; Cupertino, CA, USA
Summary:This invention describes a process of obtaining an mRNA preparation or a cDNA library from undifferentiated primate pluripotent stem (pPS) cells grown in an environment with a conditioned medium free of feeder cells. The invention relates to a system for obtaining expression libraries from pPS cells which can be maintained in vitro without requiring a layer of feeder cells to inhibit differentiation. Further disclosed are conditions that promote pPS cell growth without differentiation such as support of the culture on an extracellular matrix, and by culturing the cells in a medium conditioned by another cell type. Claims of the invention include cDNA libraries from such cultures that are devoid of transcripts of feeder cell origin, relatively uncontaminated by transcripts from differentiated cells, and can have a high proportion of full-length transcripts. Further provided are subtraction libraries which can be produced that are enriched for transcripts modulated during differentiation.
Abstract:This disclosure provides a system for obtaining expression libraries from primate pluripotent stem (pPS) cells. pPS cells can be maintained in vitro without requiring a layer of feeder cells to inhibit differentiation. The role of the feeder cells is replaced by several other culture conditions provided in a suitable combination. Conditions that promote pPS cell growth without differentiation include supporting the culture on an extracellular matrix, and culturing the cells in a medium conditioned by another cell type. The cDNA libraries from such cultures are devoid of transcripts of feeder cell origin, relatively uncontaminated by transcripts from differentiated cells, and can have a high proportion of full-length transcripts. Subtraction libraries can also be produced that are enriched for transcripts modulated during differentiation.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6667176
Application Number:US2000000688031
Date Filed:10/10/2000
Date Published:23/12/2003
Assignee:Geron Corporation, Menlo Park, CA, USA


 
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