Therapeutic serum obtained from co-cultured cells
A therapeutic serum suitable for inclusion in a cosmetic preparation may be produced by stressing a co-culture including proliferative cells. The co-culture of cells may be obtained by first establishing a monolayer of cells on a surface. After a monolayer of first culture is established, a second culture comprising more resilient and/or aggressive cells may be over-seeded on the monolayer and established. Additional cultures comprising increasingly dominant cells may then be over-seeded and established until a monolayer having the desired population of cells is obtained. The monolayer is then stressed to obtain a serum by conditioning a collection medium. The obtained serum may be combined with a suitable cosmetic base to provide a cosmetic preparation.
1. A method of producing a serum, comprising the steps of:
a) seeding a first surface with a first culture of cells;
b) providing the first culture with a first growth medium comprising nutrients and at least one growth factor;
c) growing the first culture to a monolayer of less than 100% confluence;
d) over-seeding a second culture onto the first surface and the monolayer made in step (c), said second culture comprising cells more dominant than those in the first culture;
e) growing the first and second cultures to less than 100% confluence in the presence of the first growth medium;
f) after growing the first and second cultures to less than 100% confluence, replacing the first growth medium with a collection medium, said collection medium lacking at least one of the growth factors of the first growth medium;
g) maintaining the first and second cultures for a period of time in the presence of the collection medium to produce a conditioned medium; and
h) collecting at least a portion of the conditioned medium after the period of time, wherein said conditioned medium is a therapeutic serum.
2. The method of claim 1 , further comprising growing the second culture on a second surface to less than 100% confluence before over-seeding onto the first surface.
3. The method of claim 1 , wherein the second culture is grown in the presence of a second growth medium before over-seeding onto the first surface, said second growth medium comprising nutrients and at least one growth factor.
4. The method of claim 3 , wherein the nutrients of the second growth media are provided by a basal medium.
5. The method of claim 3 , wherein the growth factors of the second growth medium comprise at least one selected from the group consisting of L-glutamine, hydrocortisone hemisuccinate, lineolic acid, lecithin, human serum albumin, basic fibroblast growth factor, epidermal growth factor, transforming growth factor, insulin, and vitamin C.
6. The method of claim 1 , wherein the nutrients of the first growth media are provided by a basal medium.
7. The method of claim 1 , wherein the growth factors of the first growth medium comprise at least one selected from the group consisting of bovine pituitary extract, L-glutamine, hydrocortisone hemisuccinate, transforming growth factor, insulin, epinephrine, and ApoTransferrin.
8. The method of claim 1 , wherein the collection medium comprises the nutrients of the first growth medium.
9. The method of claim 1 , further comprising: after collecting at least a portion of the conditioned medium, replacing the conditioned medium with the first growth medium; and
culturing the first culture and second culture in the first growth medium for a recovery period of time.
10. The method of claim 1 , further comprising: before over-seeding the second culture onto the first surface, growing the first culture to a monolayer of approximately 80 to 90% confluence; and creating voids in the monolayer of the first culture.