US4818418A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Blood partitioning method
Assignee: Becton Dickinson and Co
•
Filing: 1988-02-03
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Granted: 1989-04-04
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Expires: 2008-02-03
Abstract
Abstract The present invention is directed to a method for separating mononuclear cells from other cells in a blood sample which is more rapid than the currently used density gradient material and which provides a better separation than methods utilizing thixotropic, gel-like material. In the method, a water soluble density gradient material is placed in a centrifuged tube. A water insoluble, thixotropic, gel-like substance is also placed in the container. A blood sample is placed in the container and the container is centrifuged for a time during which various components interchange positions and the gel-like substance forms a barrier between the mononuclear cells and the other cells of the blood sample.
Inventors
- 👤 Alex M. Saunders
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 09:30
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/483; G01N33/487; G01N33/49; G01N33/491; B; B01; B01D; B01D43/00; Y; Y10; Y10T; Y10T436/00; Y10T436/25; Y10T436/25375
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
211 words
A method of separating and partitioning mononuclear cells from a blood sample, which comprises the steps of: placing a water soluble density gradient material in a centrifuge tube having a bottom closed end and positioning the water soluble density gradient material adjacent to the bottom closed end of the tube; placing a thixotropic gel-like substance in the centrifuge tube and positioning the thixotropic gel-like substance adjacent to and on top of the water soluble density gradient material, the thixotropic gel-like substance being substantially hydrophobic, and the water soluble density gradient material having a specific gravity which is higher than that of the thixotropic gel-like substance; placing a sample of blood in the centrifuge tube, the thixotropic gel-like substance acting as a partition between the water soluble density gradient material and the blood sample placed in the tube on top of the gel-like substance and acting to keep the water soluble density gradient material in place in the bottom of the tube and separated from the blood sample; and centrifuging the tube to separate the mononuclear cells from other cells of the blood sample, wherein the water soluble density gradient material and the thixotropic gel-like substance form separate barrier layers between the mononuclear cells and the other cells of the blood sample.
Claim 2
Independent Claim
211 words
A method of separating and partitioning mononuclear cells from a blood sample, which comprises the steps of: placing a water soluble density gradient material in a centrifuge tube having a bottom closed end and positioning the water soluble density gradient material adjacent to the bottom closed end of the tube; placing a thixotropic gel-like substance in the centrifuge tube and positioning the thixotropic gel-like substance adjacent to and on top of the water soluble density gradient material, the thixotropic gel-like substance being substantially hydrophobic, and the water soluble density gradient material having a specific gravity which is higher than that of the thixotropic gel-like substance; placing a sample of blood in the centrifuge tube, the thixotropic gel-like substance acting as a partition between the water soluble density gradient material and the blood sample placed in the tube on top of the gel-like substance and acting to keep the water soluble density gradient material in place in the bottom of the tube and separated from the blood sample; and centrifuging the tube to separate the mononuclear cells from other cells of the blood sample, wherein the water soluble density gradient material and the thixotropic gel-like substance form separate barrier layers between the mononuclear cells and the other cells of the blood sample.
Claim 3
Dependent Claim
30 words
A method in accordance with claim 1 wherein said tube with the blood sample contained therein is centrifuged at a force of from about 600 g to about 2000 g.
Claim 4
Dependent Claim
29 words
A method in accordance with claim 1 wherein said tube with the blood sample contained therein is centrifuged for a time of from about 5 to about 20 minutes.
Claim 5
Dependent Claim
14 words
A method in accordance with claim 1 wherein said gel-like substance is a polyester.
Claim 6
Dependent Claim
22 words
A method in accordance with claim 1 wherein said gel-like substance is a mixture of silicone fluid and a hydrophobic powered silica.
Claim 7
Dependent Claim
22 words
A method in accordance with claim 1 wherein said gel-like substance is a mixture of a hydrocarbon polymer and a powdered silica.
Claim 8
Dependent Claim
22 words
A method in accordance with claim 1 wherein said density gradient material is an aqueous solution of a polymeric saccharide and diatrizoate.
Claim 9
Dependent Claim
99 words
A method in accordance with claim 1 wherein the steps of positioning the water soluble density gradient material adjacent to the bottom closed end of the tube and positioning the thixotropic gel-like substance adjacent to and on top of the water soluble density gradient material includes the step of pre-centrifuging the tube prior to placing said blood sample within said tube to cause the water soluble density gradient material to migrate to the bottom end of the tube and to cause the thixotropic gel-like substance to form a barrier over the top of the water soluble density gradient material.
Claim 10
Dependent Claim
24 words
A method in accordance with claim 1 wherein said thixotropic, gel-like substance has a specific gravity of from about 1.07 to about 1.085 g/cc.
Claim 11
Dependent Claim
24 words
A method in accordance with claim 1 wherein said thixotropic, gel-like substance has a specific gravity of from about 1.075 to about 1.08 g/cc.
Claim 12
Dependent Claim
24 words
A method in accordance with claim 1 wherein said density gradient material has a specific gravity of from about 1.080 to about 1.100 g/cc.
Claim 13
Dependent Claim
24 words
A method in accordance with claim 1 wherein said density gradient material has a specific gravity of from about 1.085 to about 1.095 g/cc.
Claim 14
Dependent Claim
31 words
A method in accordance with claim 1 wherein said thixotropic, gel-like material has a specific gravity of 1.077 g/cc and said density gradient material has a specific gravity of 1.09 g/cc.
Claim 15
Dependent Claim
43 words
A method as defined by claim 1, wherein the thixotropic gel-like substance is selected from the group consisting of a polyester, a mixture of a silicone fluid and a hydrophobic powdered silica and a mixture of a hydrocarbon polymer and a powdered silica.
References Cited (Backward Citations)
US3852194A
Cited
US3945928A
Cited
US3920549A
Cited
US3960727A
Cited
US4101422A
Cited
US4153739A
Cited
US4350593A
Cited
US4147628A
Cited
GB2014879A
Cited
US4190535A
Cited
US4255256A
Cited
US4310430A
Cited
EP0036168A2
Cited
US4457782A
Cited
US4534798A
Cited
CA1127537A
Cited
US4417981A
Cited
US4435293A
Cited
US4436631A
Cited
US4487700A
Cited
US4751001A
Cited
US4640785A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-04-03 | 1992-10-03 | 1993-04-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-04-03 | 1996-10-03 | 1997-04-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-04-03 | 2000-10-03 | 2001-04-04 | $7700.00 | Window Closed / Lapsed |