ACTIVATED LEUKOCYTE CONDITIONED SUPERNATANT AND USES FOR WOUND HEALING
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Abstract
Disclosed are therapeutic, activated leukocyte conditioned supernatants, methods of making them, and methods of using the conditioned supernatants to repair or promote healing of wounds.
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78 Claims
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1-62. -62. (canceled)
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63. A substantially cell-free activated leukocyte-conditioned supernatant (ALCS), wherein the ALCS is made by:
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incubating human leukocytes under conditions of time and temperature to activate the leukocytes, wherein the activated leukocytes comprise activated granulocytes, activated monocytes, and activated lymphocytes; subjecting the activated leukocytes to hypo-osmotic shock; adding to the activated leukocytes subjected to the hypo-osmotic shock to a physiologically acceptable solution containing inorganic ions, organic osmolytes, or a combination thereof in an amount which restores isotonicity; separating the activated leukocytes from the resulting isotonic solution; mixing the separated activated leukocytes with a medium to form an incubation composition; incubating the incubation composition (i) at room temperature for 8 to 72 hours;
or (ii) at about 37°
C. for 3 to 5 hours; andseparating the activated leukocytes and other cellular matter from the incubation composition to produce a substantially cell-free ALCS. - View Dependent Claims (64, 65, 66, 72, 73, 74, 75, 76, 77)
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67. A method of making a substantially cell-free ALCS activated leukocyte-conditioned supernatant (ALCS), comprising:
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incubating human leukocytes under conditions of time and temperature to activate the leukocytes; subjecting the leukocytes to hypo-osmotic shock; adding to the leukocytes subjected to the hypo-osmotic shock to a physiologically acceptable solution containing inorganic ions, organic osmolytes, or a combination thereof in an amount which restores isotonicity; separating the leukocytes from the resulting isotonic solution; mixing the separated leukocytes with a medium to form an incubation composition; incubating the incubation composition (i) at room temperature for 8 to 72 hours;
or (ii) at about 37°
C. for 3 to 5 hours; andseparating the leukocytes and other cellular matter from the incubation composition to produce a substantially cell-free ALCS. - View Dependent Claims (68, 69, 70, 71, 78)
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Specification