Method for Purifying Certain Cell Populations in Blood or Bone marrow by Depleting Others
AssigneeSyngen, Inc.
InventorPhilip H. Coelho
Life of this patent
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Abstract
A method for purifying and harvesting certain cell populations in blood or bone marrow by depleting at least one of red blood cells, granulocytes, or platelets from a sample comprising blood, bone marrow, or stromal vascular fraction cells separated from adipose tissue is disclosed. The apparatus comprises a sterile, single use rigid, self-supporting cartridge within which the automated depletion, purification and harvesting of target cell populations occurs and all components may be distributed.
Claims58
A method for depleting at least one of red blood cells, granulocytes, or platelets from a sample comprising blood, bone marrow, or stromal vascular fraction cells separated from adipose tissue, the method comprising:
- a. placing a rigid cartridge within a centrifuge, said rigid cartridge comprising a rigid chamber having an end that is fluidly connected to a valve system comprising at least one valve and which is initially closed, a first rigid storage compartment, and a second rigid storage compartment;
- b. transferring said sample into said rigid chamber, said sample comprising platelets, plasma, cells of high density, and cells of low density;
- c. centrifuging said rigid cartridge such that said sample is urged toward said end by a first G force of at least 10 G and then by a second G force lower than said first G force;
- d. providing a first pathway for said cells of high density through said end and to said first rigid storage compartment;
- e. tracking the migration of said cells of high density through said end;
- and
- f. providing a second pathway for said cells of low density and an amount of platelets and plasma through said end and to said second rigid storage compartment.
A method for depleting at least one of red blood cells, granulocytes, or platelets from a sample comprising blood, bone marrow, or stromal vascular fraction cells separated from adipose tissue, the method comprising:
- a. providing:
- i. a centrifuge having an axis of rotation;
- ii. a sample comprising plasma and a first portion of high density and a remaining portion of low density;
- iii. a rigid cartridge comprising:
- 1. an internal rigid chamber having an exit port;
- 2. a first rigid storage compartment and a second rigid storage compartment;
- 3. an input port;
- and
- 4. a valve system comprising at least one valve and providing communication between said exit port and said rigid storage compartments;
- and
- iv. a sensor;
- b. placing said sample within said rigid cartridge by transferring said sample through said input port and into said rigid chamber;
- c. centrifuging said rigid cartridge such that said first portion is first urged toward said exit port by centrifugal force, directed by said valve system, and then urged toward said axis of rotation and into said first rigid storage compartment;
- and
- d. directing with said valve system an amount of said remaining portion toward said axis of rotation and into said second rigid storage compartment.
A method for depleting at least one of red blood cells, granulocytes, or platelets from a sample comprising blood, bone marrow, or stromal vascular fraction cells separated from adipose tissue, the method comprising:
- a. providing a rigid cartridge comprising:
- i. a rigid outer shell;
- ii. a generally funnel shaped internal rigid chamber having a small end comprising an output opening and a large end comprising an input opening;
- iii. a first and second rigid storage compartment initially not in fluid communication with said small end;
- iv. a first valve in communication with said output opening and said first rigid storage compartment, wherein said first valve is initially closed;
- and
- v. a second valve in communication with said output opening and said second rigid storage compartment, wherein said second valve is initially closed;
- b. providing a centrifuge configured to accept said rigid cartridge;
- c. providing a sample comprising a mixture of high density cells, low density cells, platelets and plasma;
- d. transferring said sample into said rigid cartridge via said input opening;
- e. placing said rigid cartridge in said centrifuge;
- f. applying centrifugal force to urge said sample towards said small end;
- g. stratifying said sample such that a substantial majority of said high density cells form a high density component layer and a substantial majority of said low density cells form a low density component layer;
- and
- h. opening said first valve such that said component layers migrate toward said small end and wherein said substantial majority of said high density cells are urged by centrifugal force to flow first away from said axis of rotation and then toward said axis of rotation and into said first rigid storage compartment.
A method for harvesting a substantially pure solution of at least one cell type from a sample comprising high density cells, low density cells, platelets and plasma, the method comprising:
- a. providing a rigid cartridge comprising:
- i. a generally funnel shaped internal rigid chamber having a first and second exit port, said ports initially closed;
- and
- ii. at least two rigid storage compartments;
- b. placing a biological fluid sample comprising high density cells, low density cells within said rigid chamber;
- c. centrifuging said rigid cartridge such that a substantial majority of said high density cells form a high density component layer and a substantial majority of said low density cells form a low density component layer;
- and
- d. during said centrifuging step:
- i. opening said first exit port allowing passage of a portion of said high density component layer;
- ii. closing said first port;
- and
- iii. opening said second port allowing passage of a portion of said low density component layer.
A method for harvesting mononuclear cells from a sample of blood, bone marrow, or stromal vascular fraction cells separated from adipose tissue wherein all steps occur within a single rigid cartridge, the method comprising:
- a. providing a centrifuge having an axis of rotation;
- b. providing a rigid cartridge comprising an internal rigid chamber;
- c. placing said sample into said rigid chamber, the sample comprising at least two biological components selected from the group of red blood cells, granulocytes, mononuclear cells, stem cells, platelets and plasma;
- d. inserting said rigid cartridge into said centrifuge;
- e. supplying with said centrifuge a centrifugal force to said sample, said centrifugal force:
- i. firstly displacing a majority of said red blood cells in said sample away from said axis of rotation, out of said rigid chamber, toward said axis of rotation, and into a first rigid storage compartment;
- and
- ii. secondly displacing a majority of said mononuclear cells in said sample away from said axis of rotation, out of said rigid chamber, toward said axis of rotation, and into a second rigid storage compartment.
A method for selectively depleting cells of differing densities from a sample, the method comprising:
- a. placing a rigid cartridge within a centrifuge, said rigid cartridge comprising a rigid chamber having an end fluidly connected to a valve system comprising at least one valve and which is initially closed, and at least one rigid storage compartment;
- b. putting said sample in said rigid chamber, said sample comprising cells of relatively high and low density and a fluid;
- c. centrifuging said rigid cartridge such that said sample is urged towards said end by a first G force;
- d. centrifuging said rigid cartridge such that said sample is urged towards said end by a second G force lower than said first G force and providing an open pathway through said valve system for at least a portion of said cells of relatively high density through said end and to said at least one rigid storage compartment;
- e. tracking the migration of said cells through said end;
- and
- f. closing said open pathway.
Dependencies are inferred from the claim text.