Compositions derived from stem cell released molecules and methods for formulation thereof
Compositions for use in treatment of a variety of tissue diseases include stem cells and stem cell released molecules (SRM's) suspended in an aqueous solution with a cellulosic material or other thickening agent. The stem cells and SRM's can be derived from one or more distinct cell lines. The SRM's can further include one or more mucins, cytokines, or growth factors. Exemplary formulations include stem cells and SRMs derived from epithelial stem cells, corneal limbal stem cells, and fibroblasts. Other compositions and methods for formulation thereof are described.
1. A method for forming a therapeutic composition, comprising:
culturing cells of a first cell line and a second cell line in a first and a second respective culture medium, wherein said cells secrete extracellular products into the respective culture medium so that a first conditioned medium and a second conditioned medium are respectively formed;
harvesting said first and second conditioned medium by removing cultured cells from said respective conditioned medium; and
combining said first conditioned medium and second conditioned medium to form a therapeutic composition,
wherein said first and second cell lines are an adult human adipose-derived stem cell line (ADSC), and a fetal human fibroblast cell line.
2. The method of claim 1 , wherein said first cell line is a StemPro® human adipose-derived stem cell line (ADSC)provided by Life Technologies Catalog Number R7788115.
3. The method of claim 1 , wherein said second cell line is a fetal human dermal fibroblast (HDF-f) cell line provided by ScienCell Research Laboratories Catalog Number 2300.
4. The method of claim 1 , wherein said first conditioned medium and said second conditioned medium are combined in a ratio of about 50/50 by volume.
5. The method of claim 1 , further comprising
culturing cells of a third cell line in a third culture medium, wherein said cells of said third cell line are different from those of the first and second cell lines and they secrete extracellular products into said third culture medium so that a third conditioned medium is formed; and
incorporating said third conditioned medium into said therapeutic composition,
wherein said third cell line is a human hair dermal papilla cell line (HHDPC).
6. The method of claim 5 , wherein cells of said human hair dermal papilla cell line (HHDPC) are cultured only through early passage.
7. The method of claim 5 , wherein said first, second, and third conditioned media are combined in a ratio of about 33/33/33 by volume.
8. The method of claim 1 , further comprising incorporating a thickening agent into each of said conditioned media prior to the combining step.
9. The method of claim 8 , wherein said thickening agent comprises one or more cellulosic material or polymer.
10. The method of claim 9 , wherein said thickening agent comprises one or more cellulosic material selected from the group consisting of methyl cellulose, sodium carboxymethyl cellulose, and hydroxypropylmethyl cellulose.
11. The method of claim 9 , wherein said thickening agent comprises one or more polymer selected from the group consisting of carboxyvinyl polymer, polyvinyl alcohol, polyvinyl pyrolidone, polyethylene glycol, and polysorbate 80.
12. The method of claim 1 , wherein said first cell line is a StemPro® human adipose-derived stem cell line (ADSC) provided by Life Technologies Catalog Number R87788115, and the second cell line is a fetal human dermal fibroblast (HDF-f) cell line provided by ScienCell Research Laboratories Catalog Number 2300.
13. A therapeutic composition formed by a method according to claim 1 .
14. The therapeutic composition of claim 13 , wherein said therapeutic composition is formulated for topical administration.