Abstract: This invention relates to sequential methods of cryopreserving bone marrow stromal cells that are transfected and used for gene therapy by transplantation. These methods include the following steps in various orders: obtaining the cells, expanding the cells in culture, transfecting the cells, and cryopreserving the cells. With these methods, populations of bone marrow stromal cells can be acquired that are large enough to be useful in a number of therapies. Further, these large populations can be stored for extended periods of time for immediate use when needed.
Abstract: The methods of the present invention are based on the discovery that adverse side effects (such as hemorrhage) can occur upon infusion of cells that express tissue factor. Accordingly, the methods of the invention are aimed at reducing the biological activity of tissue factor (TF) in a patient, and can be carried out by, for example: infusing fewer cells (or infuse the same number of BMSCs over a longer period of time); reducing the expression or activity of TF (within the infused cells specifically (e.g., by contacting the cells with a TF antagonist in vitro) or within the patient generally (e.g., by administering a TF antagonist to the patient); hampering the interaction of TF with factor VII(a); inhibiting the activity of the TF-factor VII(a) complex once it has formed; or inhibiting the coagulation cascade at any point downstream from formation of the complex (including inhibition of platelet activation).
Type:
Grant
Filed:
December 31, 1998
Date of Patent:
May 14, 2002
Assignee:
ALG Company
Inventors:
David R. Hurwitz, Van Cherington, Theofanis Galanopoulos, Peter H. Levine, Joel S. Greenberger
Abstract: A method of causing production and secretion into the bloodstream of a human patient of a biologically active enzyme for which the human patient suffers a deficiency; the method involves introducing into the human patient donor bone marrow stromal cells which have been transfected with a gene encoding the enzyme, so that the introduced cells can adhere to a bone cavity surface of the patient and produce and secrete the active enzyme.
Abstract: The invention features a method for the selection and expansion of bone marrow stromal cells. The method includes the steps of obtaining bone marrow stromal cells; introducing the stromal cells into a vessel pre-coated on an inner surface with a gelatin, and containing a culture medium including an acidic fibroblast growth factor ("aFGF") polypeptide; and expanding the stromal cells in the culture medium under conditions and for a time sufficient to obtain an increased number of bone marrow stromal cells. The culture medium additionally can include heparin, and the vessel additionally can be precoated with fetal bovine serum.
Abstract: The invention features a method for the selection and expansion of bone marrow stromal cells. The method includes the steps of obtaining bone marrow stromal cells; introducing the stromal cells into a vessel pre-coated on an inner surface with a gelatin, and containing a culture medium including an acidic fibroblast growth factor ("aFGF") polypeptide; and expanding the stromal cells in the culture medium under conditions and for a time sufficient to obtain an increased number of bone marrow stromal cells. The culture medium additionally can include heparin, and the vessel additionally can be precoated with fetal bovine serum.