Isolation of bone marrow fraction rich in connective tissue growth components and the use thereof to promote connective tissue formation
First Claim
1. A method for obtaining a bone marrow fraction, comprising:
- a) centrifuging a biological sample including bone marrow and whole blood to provide a separation of components of the sample based on density, said separation providing the following fractions in decreasing order of density;
(i) a fraction rich in blood cells;
(ii) a buffy coat fraction;
(iii) a platelet rich fraction; and
(iv) a platelet poor fraction; and
b) isolating the buffy coat fraction alone or in combination with all or part of the platelet rich fraction to form an isolate rich in connective tissue growth promoting components.
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Abstract
A bone marrow isolate rich in one or more connective tissue growth components, methods of forming the isolate, and methods of promoting connective tissue growth using the isolate are described. A biological sample comprising bone marrow is centrifuged to separate the sample into fractions including a fraction rich in connective tissue growth components. The fraction rich in connective tissue growth components is then isolated from the separated sample. The isolate can be used directly or combined with a carrier and implanted into a patient at a tissue (e.g., bone) defect site. The biological sample can comprise bone marrow and whole blood. The isolate can be modified (e.g., by transfection with a nucleic acid encoding an osteoinductive polypeptide operably linked to a promoter) prior to application to the tissue defect site. The isolate can be made and applied to the tissue defect site in a single procedure (i.e., intraoperatively).
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Citations
62 Claims
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1. A method for obtaining a bone marrow fraction, comprising:
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a) centrifuging a biological sample including bone marrow and whole blood to provide a separation of components of the sample based on density, said separation providing the following fractions in decreasing order of density;
(i) a fraction rich in blood cells;
(ii) a buffy coat fraction;
(iii) a platelet rich fraction; and
(iv) a platelet poor fraction; and
b) isolating the buffy coat fraction alone or in combination with all or part of the platelet rich fraction to form an isolate rich in connective tissue growth promoting components. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method for treating a patient, comprising:
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obtaining an isolate including a bone marrow fraction having components that promote connective tissue formation; and
implanting said isolate into a patient at a tissue defect site;
wherein said obtaining is performed intraoperatively with said implanting. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
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41. An implant composition, comprising:
an isolate material containing an uncultured bone marrow fraction including tissue growth promoting components, in combination with a biocompatible carrier. - View Dependent Claims (42, 43, 44, 45, 46, 47, 48)
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49. A method for treating a patient, comprising:
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providing a biological sample including tissue from a bone marrow source of the patient, said sample including tissue growth promoting components;
centrifuging the biological sample to separate the sample into fractions based on density, said fractions including a fraction rich in said tissue growth promoting components;
isolating said fraction rich in tissue growth promoting components;
implanting said fraction rich in tissue growth promoting components into the patient; and
wherein said centrifuging, and isolating are performed intraoperatively with said implanting. - View Dependent Claims (50, 52, 53, 54, 55, 56, 57)
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58. A method for obtaining an isolate material comprising tissue growth promoting components, comprising:
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harvesting tissue from a bone marrow source of a mammal, said tissue including tissue growth promoting components;
forming an admixture by combining said tissue with whole blood of the mammal;
centrifuging a biological sample including said admixture so as to separate the biological sample into fractions based on density, said fractions including a fraction rich in said tissue growth promoting components; and
isolating said fraction rich in said tissue growth promoting components.
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59. A method for obtaining a bone marrow fraction rich in connective tissue growth promoting components, the method comprising:
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centrifuging a biological sample comprising bone marrow to separate components of the sample into fractions based on density, said fractions including a fraction rich in tissue promoting components; and
isolating said fraction rich in tissue growth promoting components.
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60. A method for preparing a medical implant material for delivery, the method comprising:
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(a) centrifuging a biological sample including a bone marrow material under conditions effective to separate a fraction including bone marrow components that promote tissue growth; and
(b) loading the fraction into a device for delivering the fraction to a patient without washing the fraction.
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61. A method for preparing a medical implant material for delivery, the method comprising:
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(a) centrifuging a biological sample including a bone marrow material under conditions effective to separate a fraction including bone marrow components that promote tissue growth; and
(b) loading the fraction into a device for delivering the fraction to a patient without culturing the fraction.
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62. A medical combination comprising an isolated, uncultured bone marrow fraction enriched in components that promote tissue growth, in combination with a device for delivering the material to a patient.
Specification