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US 9,168,287 B2Granted
Activated leukocyte conditioned supernatant and uses for wound healing
AssigneeMacrocure Ltd.
InventorsMitchell Shirvan, Eilat Bain, Marina Bubis, Irene Ginis
Life of this patent
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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.
Claims12
A substantially cell-free activated leukocyte-conditioned supernatant (ALCS), wherein the ALCS is made by:
- 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 serum to form an incubation composition;
- incubating the incubation composition ii) at room temperature for 8 to 72 hours;
- or (ii) at about 37° C. for 3 to 5 hours;
- and
- separating the activated leukocytes and other cellular matter from the incubation composition to produce a substantially cell-free ALCS.
6 more elements
A method of making a substantially cell-free ALCS activated leukocyte-conditioned supernatant (ALCS), comprising:
- 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 serum 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;
- and
- separating the leukocytes and other cellular matter from the incubation composition to produce a substantially cell-free ALCS.
6 more elements
Dependencies are inferred from the claim text.