EXTRACELLULAR VESICLES DERIVED FROM CELLS CULTURED UNDER HYPOXIC CONDITIONS AND USES THEREOF
The present invention is directed to a method of producing extracellular vesicles compositions from conditioned culture medium (CCM) compositions. The method includes culturing cells under hypoxic conditions on a biocompatible surface in vitro. The culturing method produces both soluble (CCM) and non-soluble fractions. Extracellular vesicles are derived from the soluble fraction (the CCM) produced by the cells cultured under hypoxic conditions and may be used in a variety of medical and therapeutic applications.
1 . A CD9, CD 63 and/or CD81-positive extracellular vesicle (ECVs) comprising Wnt and/or follistatin proteins or a bioactive fragment thereof.
2 . The ECV of claim 1 , wherein the ECV is produced by culturing fibroblast cells under hypoxic conditions on a microcarrier bead or three dimensional surface in a suitable growth medium, under 1-5% oxygen for at least 2 weeks, thereby producing multipotent stem cells which produce and secrete ECVs into the culture medium, and collecting the ECVs.
3 . The ECV of claim 1 , wherein the Wnt protein is selected from Wnt3 a, Wnt7 a and/or Wnt7b.
4 . A method of producing an ECV comprising at least a Wnt and follistatin protein comprising culturing fibroblast cells under hypoxic conditions on a microcarrier bead or three dimensional surface in a suitable growth medium, under 1-5% oxygen for at least 2 weeks, thereby producing multipotent stem cells, wherein the multipotent stem cells produce and secrete extracellular vesicles in the culture medium.
5 . A composition comprising the extracellular vesicle of claim 1 and a carrier.
6 . The composition of claim 5 , wherein the carrier is a pharmaceutically acceptable carrier.
7 . A method for stimulating hair growth comprising administering to a subject in need thereof, an ECV of claim 1 , thereby stimulating hair growth.
8 . A method of stimulating skin rejuvenation and/or regeneration comprising administering to a subject in need thereof, an ECV of claim 1 , thereby stimulating skin rejuvenation and/or regeneration.
9 . Use of an ECV of claim 1 , for the preparation of a medicament for stimulating hair growth.
10 . Use of an ECV of claim 1 , for the preparation of a medicament for stimulating skin rejuvenation and/or regeneration.
11 . A method of treating tissue in need of repair, comprising administering to a subject in need thereof, an ECV of claim 1 , thereby repairing the tissue.
12 . The ECV of claim 1 , wherein the ECV is an exosome.