Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5081030

Title:Release of cells from affinity matrices
Inventors:Civin, Curt I.; Baltimore, MD, USA
Summary:Described herein are methods of releasing cells from cell receptor affinity complexes. The invention relates to a process for the release from the cell-receptor complex of positively selected cells in viable, fully functional condition, wherein a ligand that is involved in the particular receptor-ligand interaction that is utilized for the affinity purification is selectively attacked by one or more degradative enzymes which are specific for that ligand. Further disclosed are properties of the resulting cell suspension which can be obtained substantially free of receptor material. Multiple embodiments of the invention are provided, which include a method for positive stem cell selection via the use of anti-MY10 and immunomagnetic microspheres to isolate CD34+ marrow cells by the use of an enzyme to release micropheres from the isolated CD34+ cells, such that reproducible enzymatic cleaving of immunomagnetic microspheres from MY10+ cells may be achieved by treatment of the preparation with papain or chymopapain. Therapeutic applications are included for the release of stem cells in bone marrow transplantation. This invention represents a continuation of other applications filed by the same assignee and inventor.
Abstract:The present invention comprises a process for release from the cell-receptor complex of positively selected cells in viable, functional condition, where a ligand involved in the particular receptor-ligand interaction utilized for the affinity purification is selectively attacked by one or more degradative enzymes specific for that ligand. A resulting cell suspension can be obtained substantially free of receptor material. This invention, in one embodiment, contemplates a method for positive stem cell selection, utilizing anti-MY10 and immunomagnetic microspheres to isolate CD34-positive marrow cells and employing an enzyme to release micropheres from the isolated CD34-positive cells. Reproducible enzymatic cleaving of immunomagnetic microspheres from MY10-positive cells can be achieved by brief treatment of the preparation with papain or chymopapain. The isolated CD34-positive cells are particularly desirable for bone marrow transplantation.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5081030
Application Number:US1989000342938
Date Filed:25/04/1989
Date Published:14/01/1992
Assignee:The Johns Hopkins University, Baltimore, MD, USA


 
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