Methods and compositions for genetically modifying primate bone marrow cells
OwnerDomenico Valerio, Victor Willem Van Beusecheminventors
Life of this patent
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Abstract
A method is provided for genetically modifying primate bone marrow cells, comprising isolating bone marrow cells from a primate and, by means which enhance the local concentration of retroviral particles, contacting the isolated bone marrow cells to cells that produce a recombinant amphotropic retrovirus with a genome based on a retroviral vector that contains the genetic information to be introduced into the bone marrow cells. Recombinant amphotropic retrovirus-producing cells, suitable for use in this method also are provided, as are genetically modified primate bone marrow cells with the capacity for regeneration in vivo.
Claims45
A method for genetically modifying primate hematopoietic stem cells, said method comprising:
- combining isolated primate hematopoietic stem cells which can undergo replication with a recombinant retrovirus comprising a genome based on a retroviral vector into which a gene X is inserted, wherein said combining uses means whereby said hematopoietic stem cells and said recombinant retrovirus are brought into close physical contact, so that chance of infection of said hematopoietic stem cells by said recombinant retrovirus is significantly enhanced.
A cell which produces a recombinant retrovirus comprising a genome based on a retroviral vector into which is inserted a gene X for introduction into isolated primate bone marrow cells by the method of cocultivating isolated primate bone marrow cells with said recombinant retrovirus.
A process for producing genetically modified primate hematopoietic cells with long term repopulating capacity, said method comprising:
- collecting both adherent and nonadherent genetically modified primate bone marrow cells from a culture in which unmodified primate bone marrow cells were combined with a recombinant retrovirus under conditions whereby retroviral particle concentration in the immediate vicinity of said unmodified bone marrow cells was enhanced so as to increase efficiency of transduction of said bone marrow cells.
Genetically modified primate hematopoietic cells with long term repopulating capacity comprising:
- a plurality of adherent and nonadherent genetically modified primate bone marrow cells.
A method for genetically modifying primate hematopoietic stem cells, said method comprising combining isolated primate hematopoietic stem cells which can undergo replication in culture with a recombinant retrovirus comprising a genome based on a retroviral vector into which a gene X is inserted, wherein said combining uses means whereby said hematopoietic stem cells and said recombinant retrovims are brought into close physical contact, so that chance of infection of said hematopoietic stem cells by said recombinant retrovirus is significantly enhanced.
A cell culture comprising:
- isolated primate hematopoietic stem cells which can undergo replication and a retrovims producing cell line which produces a recombinant retrovirus comprising a genome based on a retroviral vector into which a gene X is inserted.
A method for genetically modifying primate hematopoietic stem cells, said method comprising:
- combining isolated primate hematopoietic stem cells which can undergo replication with a recombinant retrovirus comprising a genome based on a retroviral vector into which a gene X is inserted, wherein said combining uses means whereby said hematopoietic stem cells and said recombinant retrovirus are brought into close physical contact, so that chance of infection of said hematopoietic stem cells by said recombinant retrovirus is significantly enhanced.
A cell culture comprising:
- isolated primate hematopoietic stem cells which can undergo replication and a stromal cell population, wherein said stromal cell population comprises cultured cells, extracellular matrix molecules and cytokines produced by said cultured cells.
Dependencies are inferred from the claim text.