Production of neural protective and regenerative factors from stem cells and treatment of nervous system conditions therewith
View Patent ↗A method for producing stem cell conditioned media for treatment of neurological insults is provided herein. By providing for the culture of adipose stem cells, and collecting the supernatants thereof, the supernatants have been shown to effect biological activity in preventing neural death when the supernatants are administered to a patient that has or is about to be subjected to a neural insult.
1. A method of producing a stem cell conditioned media, comprising the steps of:
a. providing at least one human adipose stem cell;
b. providing at least one medium for culturing the stem cell;
c. culturing the at least one human adipose stem cell in the at least one medium;
d. switching the medium to a growth-factor free basal media for conditioning the at least one human adipose stem cell in either normoxic or hypoxic conditions for 72 hours; and
e. collecting cell supernatants effective to treat neural insults.
2. The method of claim 1 , wherein the at least one medium is EGM2MV, and the human adipose stein cells are cultured to confluence prior to switching the medium to a growth-factor free basal media.
3. The method of claim 2 , wherein the human adipose stem cell has a surface marker profile of lin-/CD45−/c-kit-/CD90+.
4. The method of claim 3 , wherein the resultant supernatant is concentrated at least 50-fold.
5. The method of claim 4 , wherein the resultant supernatant is fractioned to remove substances less than 10 kDa.
6. The method of claim 4 , wherein the resultant supernatant is fractioned to remove substances less than 30 kDa.
7. The method of claim 3 , wherein the adipose stem cells display the surface marker profile CD34+ positive.
8. The method of claim 2 , wherein the neural insults to be treated by the cell supernatants are selected from the group consisting of hypoxia-ischemia, encephalopathy, stroke neurodegenerative disorders, and chronic or acute injuries in nervous system.
9. The method of claim 2 , wherein the adipose stem cells are obtained from lipoaspirates.
10. The method of claim 2 , wherein the human adipose stem cell is harvested from an individual to be treated for a neural insult.
11. A method of producing a stem cell conditioned media, comprising the steps of:
a. providing at least one human or rat adipose stem cell harvested from an individual to be treated for a neural insult, the at least one stem cell comprising an adipose stem cell harvested from a lipoaspirate;
b. providing at least one medium for culturing the stem cell;
c. culturing the at least one human or rat adipose stem cell in the at least one medium;
d. switching the medium to a growth-factor free basal media for conditioning the at least one human or rat adipose stem cell in either normoxic or hypoxic conditions for 72 hours; and
e. collecting cell supernatants effective to treat neural insults and introducing the cell supernatants to the individual in a therapeutic dose.