Transgenic circulating endothelial cells
First Claim
1. A process for expanding the population of endothelial cells obtained from peripheral blood comprising culturing, in contact with a collagen I-coated surface, buffy coat cells obtained from peripheral mammalian blood in the presence of a cell culture medium containing an effective amount of vascular endothelial growth factor (VEGF), and which medium is free of bovine brain extract, so as to expand the population of endothelial cells in said buffy coat cells.
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Abstract
A process is provided for expanding the population of endothelial cells obtained from peripheral blood which can be transformed with a vector comprising a DNA sequence encoding a preselected bioactive polypeptide. The resulting transgenic endothelial cells are useful to biocompatibilize implantable medical devices or can be used directly, as for gene therapy.
82 Citations
43 Claims
- 1. A process for expanding the population of endothelial cells obtained from peripheral blood comprising culturing, in contact with a collagen I-coated surface, buffy coat cells obtained from peripheral mammalian blood in the presence of a cell culture medium containing an effective amount of vascular endothelial growth factor (VEGF), and which medium is free of bovine brain extract, so as to expand the population of endothelial cells in said buffy coat cells.
- 15. A transgenic mammalian endothelial cell comprising an isolated DNA sequence encoding a recombinant Factor VIII protein.
- 17. A transgenic endothelial cell which is prepared by a process comprising stably transforming a population of circulating human endothelial cells outgrown from blood with a vector comprising an isolated DNA sequence encoding a preselected protein operably linked to a promoter functional in human endothelial cells.
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33. A diagnostic method comprising:
detecting or determining whether an expanded population of endothelial cells obtained from peripheral blood of a test mammal has an acquired or genetic indication or disease relative to a control expanded population of endothelial cells obtained from a mammal not at risk of the acquired or genetic indication or disease, wherein the expanded cells are obtained by culturing, in contact with a collagen I-coated surface, mononuclear cells from a buffy coat layer obtained from the peripheral blood of the test mammal in the presence of a cell culture medium containing an effective amount of vascular endothelial growth factor (VEGF), which medium is free of bovine brain extract, so as to yield an expanded population of endothelial cells. - View Dependent Claims (34, 35, 36, 37, 38, 39, 40, 41, 42, 43)
Specification