Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5830760

Title:Creating novel hematopoietic cell lines by expressing altered retinoic acid receptors
Inventors:Tsai, Schickwann; Redmond, WA, USA
Collins, Steven J.; Seattle, WA, USA
Summary:Described herein are novel hematopoietic stem cell lines produced by stem cells that have been transfected with a nucleic acid encoding a dominant negative suppressor of the retinoic acid receptor alpha. The invention relates to methods for establishing an immortalized hematopoietic stem cell line, namely, the lympho-hematopoietic progenitor cell line, which is capable of differentiating into erythroid, myeloid, and B lymphocytic lineages. Further disclosed are methods of producing GM-CSF dependent neutrophil progenitor cell lines that are capable of differentiating into neutrophils but not into monocytes, mast cells, or basophils, via the introduction into the bone marrow, fetal spleen, fetal liver, or other hematopoietic myeloid cells of a nucleic acid encoding a dominant negative suppressor of a retinoic acid receptor-alpha and a selectable marker. Also provided are techniques for culturing the recombinant cells in a medium containing SCF or GM-CSF with agents that allow for the selective growth of the recombinant cells in combination with a level of retinoic acid of less than approximately 10-8 M to 10-9 M. Further elucidated are methods by which the addition of a retinol compound into specific cell lines induces differentiation into neutrophils. Therapeutic applications for hematopoietic reconstitution are included.
Abstract:Methods for establishing continuous SCF dependent lympho-hematopoietic progenitor cell lines capable of differentiating into erythroid, myeloid, and B lymphocytic lineages, and GM-CSF dependent neutrophil progenitor cell lines capable of differentiating into neutrophils but not into monocytes, mast cells, or basophils, by introducing into bone marrow, fetal spleen, fetal liver, or other hematopoietic myeloid cells nucleic acid encoding a dominant negative suppressor of a retinoic acid receptor-alpha and a selectable marker, and culturing the recombinant cells in culture medium containing SCF or GM-CSF, agents allowing for selective growth of the recombinant cells, and a level of retinoic acid of less than about 10.sup.-8 M to about 10.sup.-9 M in the case of establishing neutrophilic progenitor cell lines. Addition of a retinol compound induces the latter cell line to differentiate into neutrophils.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5830760
Application Number:US1996000592383
Date Filed:26/01/1996
Date Published:03/11/1998
Assignee:Fred Hutchinson Cancer Research Center, Seattle, WA, USA


 
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