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US 9,447,382 B2Granted
Generation of induced pluripotent stem cells from small volumes of peripheral blood
AssigneeCellular Dynamics International, Inc.
InventorAmanda Mack
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Abstract
Methods and compositions relating to the production of induced pluripotent stem cells (iPS cells) are disclosed. For example, induced pluripotent stem cells may be generated from peripheral blood cells, such as human blood progenitor cells, using episomal reprogramming and feeder-free or xeno-free conditions. In certain embodiments, the invention provides novel methods for improving overall reprogramming efficiency with low number of blood progenitor cells.
Claims14
A method for producing human induced pluripotent stem (iPS) cells from hematopoietic progenitor cells, the method comprising the steps of:
- a) providing a cell population of human peripheral blood cells from one or more subjects whose cells have not been mobilized with extrinsically applied granulocyte colony-stimulating factor (G-CSF) or granulocyte macrophage colony-stimulating factor (GM-CSF), the population comprising hematopoietic progenitor cells;
- b) culturing said population under feeder-free, defined expansion conditions to promote the expansion of said hematopoietic progenitor cells;
- c) introducing exogenous EBV-based episomal genetic elements into said expanded hematopoietic progenitor cells, wherein said episomal genetic elements replicate extra-chromosomally in said cells and express iPS reprogramming factors, further wherein the exogenous episomal genetic elements have one or more polycistronic cassettes.
- d) culturing said expanded hematopoietic progenitor cells in a media that is essentially free of feeder cells or feeder cell-conditioned medium, or in a xeno-free culture, thereby producing human iPS cells from said hematopoietic progenitor cells;
- and
- e) culturing said iPS cells sufficiently to remove episomal genetic elements.
3 more elements
Dependencies are inferred from the claim text.