Method for isolation and purification of microvesicles from cell culture supernatants and biological fluids

A method of treating a burn wound in a patient in need thereof, the method comprising administering a pharmaceutical composition of isolated microvesicles to the burn wound of the patient,

• wherein the isolated microvesicles are prepared comprising the steps of:
• (a) obtaining a biological fluid containing mammalian microvesicles from a mesenchymal stem cell;
• (b) precipitating the microvesicles by adding polyethylene glycol (PEG) to the biological fluid;
• (c) washing the precipitated microvesicles to remove the PEG,
• (d) isolating the microvesicles to obtain isolated microvesicles;
• and
• (e) providing a therapeutically effective amount of the precipitated isolated microvesicles as a pharmaceutical composition,
• wherein the precipitated microvesicles range in size from 2 nm to 5000 nm,
• wherein the isolated microvesicles result in increased migration of non-proliferating cultured fibroblasts in a cell migration assay when administered to the non-proliferating cultured fibroblasts, and
• wherein administering the pharmaceutical composition of isolated microvesicles to the burn wound of the patient results in in regeneration of at least one skin tissue in the burn wound.

A method of treating a burn wound in a patient in need thereof, the method comprising

• administering a pharmaceutical composition of isolated microvesicles directly to the burn wound of the patient,
• wherein the isolated microvesicles are prepared comprising the steps of:
• (a) obtaining culture medium conditioned using human bone marrow-derived mesenchymal stem cells, the culture medium containing microvesicles;
• (b) precipitating the microvesicles by adding polyethylene glycol (PEG) to the biological fluid;
• (c) washing the precipitated microvesicles to remove the PEG;
• (d) isolating the microvesicles to obtain isolated microvesicles;
• and
• (e) providing a therapeutically effective amount of the isolated microvesicles as a pharmaceutical composition,
• wherein the precipitated microvesicles range in size from 2 nm to 5000 nm,
• wherein the isolated microvesicles result in increased migration of non-proliferating cultured fibroblasts in a cell migration assay when administered to the non-proliferating cultured fibroblasts, and
• wherein administering the pharmaceutical composition of isolated microvesicles to the burn wound of the patient results in in regeneration of at least one skin tissue in the burn wound of the patient.