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US 8,529,957 B2Granted
Cell preparations for extemporaneous use, useful for healing and rejuvenation in vivo
AssigneeAntoine Turzi
InventorsAntoine Turzi, Donald Francois Du Toit
Life of this patent
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Abstract
The present invention relates to new plasma or new platelet-rich plasma preparations, new cell dissociation methods, new cell associations or compositions, a method of preparation thereof, a use thereof, devices for the preparation thereof and preparations containing such a platelet-rich plasma preparation and cell associations or compositions. Specifically, the invention provides compositions comprising plasma or platelet-rich plasma alone or in combination with cell preparations for use in tissue regeneration and bone regeneration and pain reduction.
Claims29
An isolated cell composition comprising:
- a) plasma;
- b) platelets at a concentration of at least 300×109 cells/L;
- c) white blood cells at a concentration of at least 7.0×109 cells/L;
- d) fibrinogen at a concentration of at least 3 mg/L;
- and
- e) a cell extract comprising cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- fibroblasts;
- melanocytes;
- Langerhans cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells, wherein cells are at a concentration of about 105 to about 106 cells/L or at a concentration of about 105 to about 106 cells/ml of plasma or enriched plasma;
- and wherein the erythrocyte concentration in said isolated cell composition is less than 0.6×1012 cells/L.
28 more elements
A process for the preparation of a cell composition, comprising the steps of:
- a) centrifuging whole blood in a separator tube selected from:
- a glass separator tube containing a polyester-based thixotropic gel and a buffered sodium citrate solution at 0.10 M;
- or
- a polyethylene terephthalate separator tube containing a thixotropic gel formed by a polymer mixture and an anhydrous sodium citrate at 3.5 mg/mL;
- wherein the centrifugation step is performed at a force of about 1500 g up to about 2000 g for a sufficient length of time to form a barrier between full plasma containing platelets, lymphocytes and monocytes and a pellet containing the erythrocytes;
- b) optionally separating enriched platelet rich plasma from full plasma by removing about half of the supernatant formed during the centrifuging step, said removed supernatant containing platelet poor plasma, wherein the separation is made by collecting the supernatant from atop of said barrier;
- and wherein the enriched plasma is enriched in leucocytes, thrombocytes and adhesion proteins as compared to native whole blood;
- c) re-suspending the enriched platelet rich plasma or the full plasma to form a platelet concentrate;
- d) providing a cell extract comprising cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- cornea keratinocytes;
- dermal cells;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells, gingival cells and pancreas islet cells;
- and
- e) admixing the platelet concentrate obtained under step c) with the cell extract of step d).
31 more elements
A process for the preparation of a cell composition, comprising the steps of:
- a) centrifuging whole blood in a separator tube selected from:
- a glass separator tube containing a polyester based thixotropic gel and a buffered sodium citrate solution at 0.10 M;
- or
- a polyethylene terephthalate separator tube containing a thixotropic gel formed by a polymer mixture and an anhydrous sodium citrate at 3.5 mg/mL;
- wherein the centrifugation step is performed at a force of about 1500g up to about 2000g for a sufficient length of time to form a barrier between full plasma containing platelets, lymphocytes and monocytes and a pellet containing the erythrocytes;
- b) optionally separating enriched platelet rich plasma from full plasma by removing about half of the supernatant formed during the centrifuging step, said removed supernatant containing platelet poor plasma, wherein the separation is made by collecting the supernatant from atop of said barrier;
- and wherein the enriched plasma is enriched in leucocytes, thrombocytes and adhesion proteins as compared to native whole blood;
- c) re-suspending the enriched platelet rich plasma or the full plasma to form a platelet concentrate;
- d) providing a cell extract comprising cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells;
- and
- e) admixing the platelet concentrate obtained under step c) with the cell extract of step d), wherein the cell extract is obtained by a process comprising the steps of:
- i) providing or obtaining cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells;
- ii) optionally culturing the cells of step i);
- iii) optionally harvesting the cells of step ii) if said cells are cultured;
- and
- iv) re-suspending the cultured cells obtained under step i) or step ii) or step iii) in platelet concentrate, said platelet concentrate comprising i) plasma;
- ii) platelets at a concentration of at least 300×109 cells/L;
- iii) white blood cells at a concentration of at least 7.0×109 cells/L;
- and iv) fibrinogen at a concentration of at least 3 mg/L;
- and wherein the erythrocyte concentration in said platelet concentrate is less than 0.6×1012 cells/L.
67 more elements
A process for the preparation of a cell composition comprising:
- a) providing a platelet concentrate comprising i) plasma;
- ii) platelets at a concentration of at least 300×109 cells/L;
- iii) white blood cells at a concentration of at least 7.0×109 cells/L;
- and iv) fibrinogen at a concentration of at least 3 mg/L;
- and wherein the erythrocyte concentration is less than 0.6×1012 cells/L;
- and
- b) admixing to said platelet concentrate a cell extract comprising cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- fibroblasts;
- melanocytes;
- Langerhans cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells:
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells;
- and pancreas islet cells.
29 more elements
A process for the preparation of a cell composition, comprising the steps of:
- a) providing cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells;
- i) culturing the cells in a platelet concentrate composition, wherein the final concentration of platelet concentrate is between about 5% and about 40% of the volume of the culture medium;
- ii) optionally harvesting the cells;
- and
- iii) optionally re-suspending the cultured cells obtained under step i) or ii) into a platelet concentrate composition,
- wherein said platelet concentrate composition comprises i) plasma;
- ii) platelets at a concentration of at least 300×109 cells/L;
- iii) white blood cells at a concentration of at least 7.0×109 cells/L;
- and iv) fibrinogen at a concentration of at least 3 mg/L;
- and wherein the erythrocyte concentration is less than 0.6×1012 cells/L.
32 more elements
A process for the preparation of a cell composition, comprising the steps of:
- i) providing or obtaining cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells;
- ii) optionally culturing the cells of step i);
- iii) optionally harvesting the cells of step ii) if said cells are cultured;
- and
- iv) re-suspending the cultured cells obtained under step i) or step ii) or step iii) in platelet concentrate, said platelet concentrate comprising i) plasma;
- ii) platelets at a concentration of at least 300×109 cells/L;
- iii) white blood cells at a concentration of at least 7.0×109 cells/L;
- and iv) fibrinogen at a concentration of at least 3 mg/L;
- and wherein the erythrocyte concentration in said platelet concentrate is less than 0.6×1012 cells/L.
31 more elements
A process for the preparation of a cell composition comprising:
- a) providing a platelet concentrate comprising i) plasma;
- ii) platelets at a concentration of at least 300×109 cells/L;
- iii) white blood cells at a concentration of at least 7.0×109 cells/L;
- and iv) fibrinogen at a concentration of at least 3 mg/L;
- and wherein the erythrocyte concentration is less than 0.6×1012 cells/L;
- and
- b) admixing to said platelet concentrate a cell extract comprising cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells;
- and pancreas islet cells,
- wherein said cell extract is obtained by a process comprising the steps of:
- i) providing or obtaining cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells;
- ii) optionally culturing the cells of step i);
- iii) optionally harvesting the cells of step ii) if said cells are cultured;
- and
- iv) re-suspending the cultured cells obtained under step i) or step ii) or step iii) in platelet concentrate, said platelet concentrate comprising i) plasma;
- ii) platelets at a concentration of at least 300×109 cells/L;
- iii) white blood cells at a concentration of at least 7.0×109 cells/L;
- and iv) fibrinogen at a concentration of at least 3 mg/L;
- and wherein the erythrocyte concentration in said platelet concentrate is less than 0.6×1012 cells/L.
65 more elements
A process for the preparation of a cell composition, comprising the steps of:
- a) centrifuging whole blood in a separator tube selected from:
- a glass separator tube containing a polyester based thixotropic gel and a buffered sodium citrate solution at 0.10 M;
- or
- a polyethylene terephthalate separator tube containing a thixotropic gel formed by a polymer mixture and an anhydrous sodium citrate at 3.5 mg/mL;
- wherein the centrifugation step is performed at a force of about 1500g up to about 2000g for a sufficient length of time to form a barrier between full plasma containing platelets, lymphocytes and monocytes and a pellet containing the erythrocytes;
- b) optionally separating enriched platelet rich plasma from full plasma by removing about half of the supernatant formed during the centrifuging step, said removed supernatant containing platelet poor plasma, wherein the separation is made by collecting the supernatant from atop of said barrier;
- and wherein the enriched plasma is enriched in leucocytes, thrombocytes and adhesion proteins as compared to native whole blood;
- c) obtaining the enriched platelet rich plasma to form a platelet concentrate;
- d) providing a cell extract comprising cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells;
- and
- e) admixing the platelet concentrate obtained under step c) with the cell extract of step d),
- wherein said cell extract provided in step d) is obtained by a process comprising the steps of:
- i) providing or obtaining cells selected from the group consisting of adipocytes;
- adipose stem cells;
- fat cells;
- corneal cells;
- corneal limbal stem cells;
- corneal keratinocytes;
- dermal cells;
- keratinocytes;
- fibroblasts;
- melanocytes;
- Langerhan's cells;
- bone marrow cells;
- muscle cells;
- satellite stem cells;
- myoblast progenitor stem cells;
- osteoblasts;
- chondrocytes;
- periosteal membrane cells;
- umbilical cord stem cells;
- stem cells;
- Schwann cells;
- cartilage cells;
- ligament cells;
- tendon cells;
- connective tissue cells;
- gingival cells and pancreas islet cells;
- ii) optionally culturing the cells of step i);
- iii) optionally harvesting the cells of step ii) if said cells are cultured;
- and
- iv) re-suspending the cultured cells obtained under step i) or step ii) or step iii) in platelet concentrate, said platelet concentrate comprising i) plasma;
- ii) platelets at a concentration of at least 300×109 cells/L;
- iii) white blood cells at a concentration of at least 7.0×109 cells/L;
- and iv) fibrinogen at a concentration of at least 3 mg/L;
- and wherein the erythrocyte concentration in said platelet concentrate is less than 0.6×1012 cells/L.
68 more elements
Dependencies are inferred from the claim text.