Generating a mucin-producing cell from an umbilical cord amniotic membrane epithelial stem cell
The present invention relates to the generation of a mucin-producing cell using stem/progenitor cells obtained from the amniotic membrane of umbilical cord and therapeutic uses of such mucin-producing cells.
1. A method of generating a mucin-producing cell comprising:
placing umbilical cord amniotic membrane epithelial stem cells (UCEC) in a container;
culturing the UCEC in the container in a growth medium adapted for the cultivation of a mucin-producing cell through the addition of a growth factor selected from the group consisting of epidermal growth factor (EGF), insulin-like growth factor-1, platelet-derived growth factor-BB (PDGFb), transforming growth factor-β, keratinocyte growth factor (KGF), TGF-αand amphiregulin and combinations thereof; and
permitting the UCEC to differentiate into mucin producing cells as indicated by observing mucin in the culture medium.
2. A method according to claim 1 , wherein said growth medium adapted for the cultivation of mucin-producing cells further comprises insulin, transferrin, and selenous acid.
3. A method according to claim 2 , wherein said growth medium adapted for the cultivation of mucin-producing cells comprises a medium selected from the group consisting of KGM®-Keratinocyte Medium (Cambrex), MEGM™-Mammary Epithelial Cell Medium (Cambrex), EpiLife® medium (Cascade Biologics), Green's Medium, CMRL1066 (Mediatech, Inc.), M171 (Cascade Biologics), L-15 medium, Dulbecco's Modified Eagle Medium (DMEM), DMEM-F12 and RPMI-1640 media.
4. A method according to claim 1 , wherein said medium adapted for the cultivation of a mucin-producing cell comprises a growth medium for the cultivation of mucin-producing cells, epidermal growth factor (EGF), insulin, transferrin and selenous acid.
5. A method according to claim 4 , wherein said medium adapted for the cultivation of a mucin-producing cell comprises about 98,8 to about 99,4% (v/v) of a growth medium for the cultivation of mucin-producing cells, about 0,2 to about 0,4% (v/v) insulin, about 0,2 to about 0,4% (v/v) transferrin, about 0,2 to about 0,4% (v/v) selenous acid and about 10 ng/ml epidermal growth factor (EGF).
6. A method according to claim 5 , wherein said medium adapted for the cultivation of a mucin-producing cell comprises about 98,8 to about 99,4% (v/v) CMRL1066 (Mediatech, Inc.), about 0,2 to about 0,4% (v/v) insulin, about 0,2 to about 0,4% (v/v) transferrin, about 0,2 to about 0,4% (v/v) selenous acid and about 10 ng/ml epidermal growth factor (EGF).
7. A method according to claim 6 , wherein said medium adapted for the cultivation of a mucin-producing cell comprises about 98,8to about 99,4% (v/v) M171 (Cascade Biologics), about 0,2 to about 0,4% (v/v) insulin, about 0,2 to about 0,4% (v/v) transferrin, about 0,2 to about 0,4% (v/v) selenous acid and about 10 ng/ml epidermal growth factor (EGF).