Muscle derived cells for the treatment of gastro-esophageal pathologies and methods of making and using the same
The present invention provides muscle-derived progenitor cells that show long-term survival following transplantation into body tissues and which can augment soft tissue following introduction (e.g. via injection, transplantation, or implantation) into a site of soft tissue. Also provided are methods of isolating muscle-derived progenitor cells, and methods of genetically modifying the cells for gene transfer therapy. The invention further provides methods of using compositions comprising muscle-derived progenitor cells for the augmentation and bulking of mammalian, including human, soft tissues in the treatment of various cosmetic or functional conditions, including malformation, injury, weakness, disease, or dysfunction. In particular, the present invention provides treatments and amelioration of symptoms for gastro-esophageal pathologies like gastro-esophageal reflux.
1. A method of increasing lower esophageal sphincter pressure in a mammalian subject at risk of developing gastro-esophageal reflex disease, the method comprising:
(a) isolating skeletal muscle cells from the mammalian subject;
(b) cooling the cells to a temperature lower than 10° C. and storing the cells for 1-7 days;
(c) suspending the mammalian skeletal muscle cells in media in a first cell culture container for between 30 and 120 minutes;
(d) decanting the media from the first cell culture container to a second cell culture container;
(e) allowing the remaining cells in the media to attach to the walls of the second cell culture container;
(f) isolating the cells from the walls of the second cell culture container, wherein the isolated cells are muscle-derived progenitor cells (MDCs);
(g) culturing the isolated MDCs to expand their number;
(h) freezing the MDCs to a temperature below −30° C.;
(i) thawing the MDCs; and
(j) administering the MDCs to the esophagus of the mammalian subject;
thereby increasing lower esophageal sphincter pressure by at least about 50% in the mammalian subject.
2. The method of claim 1 , wherein the skeletal muscle cells are isolated from the mammalian subject before a gastro-esophageal reflux disease begins in the mammalian subject.
3. The method of claim 1 , wherein the increase in lower esophageal sphincter pressure is increased by at least about 100%.
4. The method of claim 1 , wherein the MDCs are administered by injecting them into the esophagus.
5. The method of claim 1 , wherein the administering step involves injecting the MDCs into the lower esophageal sphincter.
6. The method of claim 1 , wherein the mammal is a human.
7. A method of increasing lower esophageal sphincter pressure in a mammalian subject comprising:
(a) isolating skeletal muscle cells from the mammalian subject,
(b) suspending the mammalian skeletal muscle cells in media in a first cell culture container for between 30 and 120 minutes;
(c) decanting the media from the first cell culture container to a second cell culture container;
(d) allowing the remaining cells in the media to attach to the walls of the second cell culture container;
(e) isolating the cells from the walls of the second cell culture container, wherein the isolated cells are muscle-derived progenitor cells (MDCs); and
(f) administering the MDCs to the esophagus of the mammalian subject;
thereby increasing lower esophageal sphincter pressure in the mammalian subject.
8. The method of claim 7 , wherein the increase in lower esophageal sphincter pressure is increased by at least about 100%.
9. The method of claim 7 , wherein the MDCs are administered by injecting them into the esophagus.
10. The method of claim 9 , wherein the MDCs are injected into the lower esophageal sphincter.
11. The method of claim 7 , wherein the mammal is a human.
12. The method of claim 7 , wherein the MDCs are cultured to expand their number before being administered to the esophagus of the mammalian subject.