Closed system separation of adherent bone marrow stem cells for regenerative medicine applications
First Claim
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1. A method for isolating and processing bone marrow derived stem cells comprising the steps of:
- (a) collecting a biological sample comprising adherent bone marrow stem cells;
(b) transferring the biological sample to a first closed receptacle containing an anticoagulant;
(c) adding to the first closed receptacle an amount of plasma blood volume expander effective to separate red blood cells from the biological sample;
(d) transferring the biological sample from the first closed receptacle to a second closed receptacle through interconnected sterile tubing, the second closed receptacle having an interior surface coated with a cell-adherent surface-layer and containing a growth medium;
(e) incubating the adherent bone marrow stem cells in said second closed receptacle for in between about two hours and about five days, wherein the adherent bone marrow stem cells adhere to the cell-adherent surface-layer, forming an adherent bone marrow stem cell layer;
(f) washing the adherent bone marrow stem cell layer, wherein any non-adherent bone marrow stem cells are removed from the adherent bone marrow stem cell layer; and
(g) collecting said adherent bone marrow stem cell layer to create collected bone marrow stem cells.
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Abstract
A method for isolating and processing bone marrow derived stem cells, including the steps of: (a) collecting a biological sample containing adherent bone marrow stem cells in a receptacle with interior walls coated with a cell-adherent substrate; (b) incubating the bone marrow cells on the adherent substrate so that a layer of adherent bone marrow stem cells adheres to the substrate; (c) washing any non-adherent cells from the substrate; and (d) collecting the bone marrow stem cell layer. Isolation kits and use of bone marrow cells harvested for cell therapies are also described.
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Citations
14 Claims
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1. A method for isolating and processing bone marrow derived stem cells comprising the steps of:
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(a) collecting a biological sample comprising adherent bone marrow stem cells; (b) transferring the biological sample to a first closed receptacle containing an anticoagulant; (c) adding to the first closed receptacle an amount of plasma blood volume expander effective to separate red blood cells from the biological sample; (d) transferring the biological sample from the first closed receptacle to a second closed receptacle through interconnected sterile tubing, the second closed receptacle having an interior surface coated with a cell-adherent surface-layer and containing a growth medium; (e) incubating the adherent bone marrow stem cells in said second closed receptacle for in between about two hours and about five days, wherein the adherent bone marrow stem cells adhere to the cell-adherent surface-layer, forming an adherent bone marrow stem cell layer; (f) washing the adherent bone marrow stem cell layer, wherein any non-adherent bone marrow stem cells are removed from the adherent bone marrow stem cell layer; and (g) collecting said adherent bone marrow stem cell layer to create collected bone marrow stem cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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Specification