Continuous selective clonogenic expansion of relatively undifferentiated cells
First Claim
1. A method of selective clonogenic expansion of a predetermined population of cells, said method comprising:
- (a) providing a chamber having an inlet, an outlet, and a passage for fluid flow from the inlet to the outlet, and, disposed in the chamber, a solid support capable of supporting a plurality of living cells adhered thereto, wherein said support comprises a plurality of selective binding molecules with specific affinity for said predetermined population of cells;
(b) causing fluid containing a plurality of cells of said cell population to flow through said chamber and selecting the flow rate of said fluid through said chamber so that a portion of said cells bind to said binding sites while unwanted cells are removed; and
(c) causing fluid containing nutrients to flow through said chamber to cause said bound cells to divide.
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Accused Products
Abstract
A system is provided for selective clonogenic expansion of relatively undifferentiated cells, including (a) a tube containing a plurality of beads of a size which permits a plurality of the undifferentiated cells to grow thereon, the beads bearing on their surfaces a selective binding molecule which binds to a surface antigen present on the relatively undifferentiated cells, wherein the antigen is not present on the surfaces of the relatively differentiated cells; (b) means for continuously providing nutrients to the relatively undifferentiated cells growing on the beads, wherein the nutrients are delivered via a fluid which flows through the tube and past the beads; and (c) means for continuously harvesting the relatively undifferentiated cells downstream of the beads.
56 Citations
26 Claims
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1. A method of selective clonogenic expansion of a predetermined population of cells, said method comprising:
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(a) providing a chamber having an inlet, an outlet, and a passage for fluid flow from the inlet to the outlet, and, disposed in the chamber, a solid support capable of supporting a plurality of living cells adhered thereto, wherein said support comprises a plurality of selective binding molecules with specific affinity for said predetermined population of cells; (b) causing fluid containing a plurality of cells of said cell population to flow through said chamber and selecting the flow rate of said fluid through said chamber so that a portion of said cells bind to said binding sites while unwanted cells are removed; and (c) causing fluid containing nutrients to flow through said chamber to cause said bound cells to divide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 26)
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21. A method of making a bioreactor for the expansion and harvesting of a predetermined population of cells, said method comprising:
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(a) providing a chamber having an inlet, an outlet, and a passage for fluid flow from the inlet to the outlet, and, disposed in the chamber, a solid support capable of supporting a plurality of living cells adhered thereto, wherein said support comprises a plurality of binding sites with specific affinity for said predetermined population of cells; (b) causing fluid containing a plurality of cells of said cell population to flow through said chamber, at a rate selected so that a portion of said cells bind to said binding sites while unwanted cells are removed; and (c) freezing said chamber to preserve said cells in a potentially viable state.
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22. A method of preserving and expanding a predetermined cell population comprising the steps of:
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(a) providing a chamber having an inlet, an outlet, and a passage for fluid flow from the inlet to the outlet, and, disposed in the chamber, a solid support capable of supporting plurality of living cells adhered thereto, wherein said support comprises a plurality of binding sites with specific affinity for said predetermined population of cells; (b) causing fluid containing a plurality of cells of said cell population to flow through said chamber, so that a portion of said cells bind to said binding sites; and (c) freezing said chamber to preserve said cells in potentially viable state; (d) thawing said chamber; (e) causing fluid containing nutrients to flow through said chamber to cause said bound cells to divide; and (f) continuously harvesting cells of said predetermined population as they exit said chamber with said fluid.
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23. A method of obtaining a desired population of cells comprising:
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(a) providing a sample containing a first population of cells which cells are the progenitor to the desired population of cells; (b) passing a portion of the sample through a chamber containing a solid support capable of supporting a plurality of living cells adhered thereto, wherein said support comprises a plurality of binding sites with specific affinity for said first population of cells so that a portion of said cells bind to said binding sites; (c) causing fluid containing nutrients to flow through said chamber at a rate selected to cause said bound cells to proliferate, differentiating to form said desired population of cells, and said desired population of cells to be released from the solid support; and (d) continuously harvesting said desired population of cells as they are released from the solid support.
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24. A method of obtaining a desired population of cells comprising:
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(a) providing a sample containing relatively undifferentiated cells; (b) passing a portion of the sample through a chamber containing a solid support capable of supporting a plurality of living cells adhered thereto, wherein said support comprises a plurality of binding sites with specific affinity for said relatively undifferentiated cells so that a portion of said cells bind to said binding sites; (c) causing fluid containing nutrients to flow through said chamber at a rate selected to cause said bound cells to proliferate and said desired population of cells to be released from the solid support, said fluid containing one or more growth factors selected to cause said cells to differentiate to form the desired population of cells; (d) continuously harvesting said desired population of cells as they exit said chamber with said fluid.
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Specification