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Stem Cell Related Patent Number US6849454

Title:Highly efficient gene transfer into human repopulating stem cells by RD114 pseudotyped retroviral vector particles
Inventors:Kelly, Patrick F.; Cincinatti, OH, USA
Vanin, Elio F.; Houston, TX, USA
Summary:Described herein are methods of highly efficient gene transfer into stem cells, particularly human hematopoietic stem cells, via contacting of the cells with viral particles that have been pseudotyped with feline endogenous retrovirus envelope protein. The invention relates to a method for efficiently introducing exogenous genes into stem cells, particularly human stem cells, optionally including the proliferation of target cells by pre-stimulation with cytokines and/or growth factors, followed by incubation of the cells with RD114-pseudotyped vector particles. In one embodiment of the invention, the vector particles are retronectin-immobilized or ultracentrifugation-concentrated retroviral vector particles pseudotyped with the feline endogenous retrovirus (RD114) envelope protein. Further disclosed are therapeutic applications such as somatic gene therapy, involving the introduction of a gene of interest contained within the retroviral genome into human repopulating stem cells followed by the introduction of these cells into a human host. Also provided is a method for monitoring the efficiency of the stem cell-mediated gene transfer based on detecting the presence of the genes, or its expression products, of the retroviral vector in various stem cell-derived lineages of the host.
Abstract:The present invention pertains to a method for efficiently introducing exogenous genes into stem cells, particularly human stem cells. The method optionally includes the steps of inducing the proliferation of target cells by pre-stimulation with cytokines and/or growth factors, followed by incubating these cells with RD114-pseudotyped vector particles. In a specific embodiment, the vector particles are retronectin-immobilized or ultracentrifugation-concentrated retroviral vector particles pseudotyped with the feline endogenous retrovirus (RD114) envelope protein. The present invention further discloses a method for somatic gene therapy, which can be used for various therapeutic applications and involves introducing a gene of interest contained within the retroviral genome into human repopulating stem cells followed by introducing these cells into a human host. Finally, the present invention discloses a method for monitoring the efficiency of the stem cell-mediated gene transfer based on detecting the presence of the genes (or the expression products) of the retroviral vector in various stem cell-derived lineages of the host.
US Patent Website:Click Here for Full Text of Patent
Title Number:US6849454
Application Number:US2001000801302
Date Filed:07/03/2001
Date Published:01/02/2005
Assignee:St. Jude Children's Research Hospital, Memphis, TN, USA


 
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