US5409813A
Expired / Lapsed
US
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Method for mammalian cell separation from a mixture of cell populations
Assignee: Systemix Inc
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Filing: 1993-09-30
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Granted: 1995-04-25
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Expires: 2013-09-30
Abstract
Abstract The invention provides a method for both the positive and negative selection of at least one mammalian cell population from a mixture of cell populations utilizing a magnetically stabilized fluidized bed. One desirable application of this method is the separation and purification of mammalian hematopoietic cells. Target cell populations include human stem cells.
Inventors
- 👤 Richard M. Schwartz
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 14:59
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/56966; B; B03; B03C; B03C1/00; B03C1/32; C; C12; C12N; C12N5/00; C12N5/0081; G01N33/543; G01N33/54313; G01N33/54326; G01N33/54333; C12M; C12M47/00; C12M47/04
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
109 words
A method for selectively separating at least one mammalian target cell population from a suspension of a mixture of cell populations wherein the suspension contains the target cell population and at least one nontarget cell population, said method comprising the steps of: fluidizing a column containing a bed of magnetizable particles with a first solution, said particles comprising a substance having affinity for said target cell population; stabilizing said fluidized bed of magnetizable particles with a radially and axially substantially uniform magnetic field; passing said suspension through said fluidized, magnetically stabilized bed at a velocity whereby said particles bind said target cell population; and collecting said target cell population.
Claim 2
Independent Claim
109 words
A method for selectively separating at least one mammalian target cell population from a suspension of a mixture of cell populations wherein the suspension contains the target cell population and at least one nontarget cell population, said method comprising the steps of: fluidizing a column containing a bed of magnetizable particles with a first solution, said particles comprising a substance having affinity for said target cell population; stabilizing said fluidized bed of magnetizable particles with a radially and axially substantially uniform magnetic field; passing said suspension through said fluidized, magnetically stabilized bed at a velocity whereby said particles bind said target cell population; and collecting said target cell population.
Claim 3
Dependent Claim
45 words
The method according to claim 1 wherein said mixture of cell populations are selected from the group consisting of fetal bone marrow, adult bone marrow, peripheral blood, fractions of peripheral blood, fetal liver, adult liver, fetal kidney, adult kidney, fetal pancreas, and adult pancreas cells.
Claim 4
Dependent Claim
18 words
The method according to claim 1 wherein said mixture of cell populations is derived from a mammalian organ.
Claim 5
Dependent Claim
16 words
The method according to claim 1 wherein said target cell population comprises human hematopoietic stem cells.
Claim 6
Dependent Claim
16 words
The method according to claim 1 wherein said target cell population comprises human hematopoietic progenitor cells.
Claim 7
Dependent Claim
18 words
The method according to claim 1 wherein said target cell population possesses hematopoietic, lymphoid, myeloid, or erythroid characteristics.
Claim 8
Dependent Claim
26 words
The method according to claim 1 wherein said collecting step further comprises the step of releasing said target cell population from said particles by physical means.
Claim 9
Dependent Claim
29 words
The method according to claim 7 wherein said releasing comprises decreasing said magnetic field to allow Brownian motion of said particles to separate the bound cells from said particles.
Claim 10
Dependent Claim
39 words
The method according to claim 7 wherein said releasing comprises passing a second solution through said bed of particles and increasing the velocity of said second solution to permit shear forces to separate the bound cells from said particles.
Claim 11
Dependent Claim
25 words
The method .according to claim 9 wherein said physical releasing further comprises decreasing said magnetic field to allow radial and axial movement of said particles.
Claim 12
Dependent Claim
26 words
The method according to claim 1 wherein said collecting step further comprises the step of releasing said target cell population from said particles by chemical means.
Claim 13
Dependent Claim
26 words
The method according to claim 11 wherein said releasing comprises passing a second solution having a pH different from said suspension through said bed of particles.
Claim 14
Dependent Claim
23 words
The method according to claim 11 wherein said chemical releasing comprises competitive inhibition binding with molecules which compete with said target cell population.
Claim 15
Dependent Claim
27 words
The method according to claim 1 wherein said magnetizable particles are in the range of 50 to 1000 microns and have a density less than 5 g/ml.
Claim 16
Dependent Claim
27 words
The method according to claim 14 wherein said magnetizable particles are in the range of 50 to 300 microns and have a density less than 2 g/ml.
Claim 17
Independent Claim
118 words
A method for selectively separating at least one target mammalian cell population from a suspension of a mixture of cell populations wherein the suspension contains: the target cell population and at least one nontarget cell population, said method comprising the steps of: fluidizing a column containing a bed of magnetizable particles with a first solution, said particles comprising a substance having affinity for the nontarget cell populations; stabilizing said fluidized bed of magnetizable particles with a radially and axially substantially uniform magnetic field; passing said suspension through said fluidized, magnetically stabilized bed of particles at a velocity whereby said particles bind at least one nontarget cell population; and collecting said target cell population which passes through said bed.
Claim 18
Independent Claim
118 words
A method for selectively separating at least one target mammalian cell population from a suspension of a mixture of cell populations wherein the suspension contains: the target cell population and at least one nontarget cell population, said method comprising the steps of: fluidizing a column containing a bed of magnetizable particles with a first solution, said particles comprising a substance having affinity for the nontarget cell populations; stabilizing said fluidized bed of magnetizable particles with a radially and axially substantially uniform magnetic field; passing said suspension through said fluidized, magnetically stabilized bed of particles at a velocity whereby said particles bind at least one nontarget cell population; and collecting said target cell population which passes through said bed.
Claim 19
Dependent Claim
45 words
The method according to claim 16 wherein said mixture of cell populations is selected from the group consisting of fetal bone marrow, adult bone marrow, peripheral blood, fractions of peripheral blood, fetal liver, adult liver, fetal kidney, adult kidney, fetal pancreas, and adult pancreas cells.
Claim 20
Dependent Claim
18 words
The method according to claim 16 wherein said mixture of cell populations is derived from a mammalian organ.
Claim 21
Dependent Claim
16 words
The method according to claim 16 wherein said target cell population comprises human hematopoietic stem cells.
Claim 22
Dependent Claim
16 words
The method according to claim 16 wherein said target cell population comprises human hematopoietic progenitor cells.
Claim 23
Dependent Claim
18 words
The method according to claim 16 wherein said target cell population possesses hematopoietic, lymphoid, myeloid, or erythroid characteristics.
Claim 24
Dependent Claim
27 words
The method according to claim 16 wherein said magnetizable particles are in the range of 50 to 1000 microns and have a density less than 5 g/ml.
Claim 25
Dependent Claim
27 words
The method according to claim 22 wherein said magnetizable particles are in the range of 50 to 300 microns and have a density less than 2 g/ml.
Claim 26
Independent Claim
119 words
A method for the selective enrichment of at least one mammalian target cell population from a suspension of a mixture of cell populations wherein said suspension contains said target cell population and at least one nontarget cell population, said method comprising the steps of: fluidizing a column containing a bed of magnetizable particles with a first solution, said particles comprising an antibody having affinity for a specific population of mammalian cells; stabilizing said fluidized bed of magnetizable particles with a radially and axially substantially uniform magnetic field; and passing the suspension through said fluidized, magnetically stabilized bed at a velocity at which at least one specific population of cells binds to said particles thereby enriching the target cell population.
Claim 27
Independent Claim
119 words
A method for the selective enrichment of at least one mammalian target cell population from a suspension of a mixture of cell populations wherein said suspension contains said target cell population and at least one nontarget cell population, said method comprising the steps of: fluidizing a column containing a bed of magnetizable particles with a first solution, said particles comprising an antibody having affinity for a specific population of mammalian cells; stabilizing said fluidized bed of magnetizable particles with a radially and axially substantially uniform magnetic field; and passing the suspension through said fluidized, magnetically stabilized bed at a velocity at which at least one specific population of cells binds to said particles thereby enriching the target cell population.
Claim 28
Dependent Claim
33 words
The method according to claim 24 wherein said particles have affinity for said target cell population and the method further comprises the step of collecting said target cell population bound to said particles.
Claim 29
Dependent Claim
36 words
The method according to claim 24 wherein said particles have affinity for said nontarget cell population and the method further comprises the step of collecting said target cells which pass through said fluidized, magnetically stabilized bed.
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