Soft tissue and bone augmentation and bulking utilizing muscle-derived progenitor cells, compositions and treatments thereof
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 for dermatological conditions, gastroesophageal reflux, vesico-ureteral reflux, urinary incontinence, fecal incontinence, heart failure, and myocardial infarction.
1. A method of augmenting or bulking the anal sphincter soft-tissue in a recipient, comprising: introducing an isolated population of cultured cells enriched in viable, desmin-expressing, skeletal muscle-derived myoblasts into the anal sphincter soft-tissue of the recipient in an amount sufficient for the myoblasts to augment or bulk the anal sphincter soft-tissue; wherein the skeletal muscle-derived myoblasts are introduced in a composition comprising a physiologically acceptable medium, wherein the skeletal muscle-derived myoblasts are isolated using a method comprising:
(a) plating a suspension of skeletal muscle cells from skeletal muscle tissue in a first container containing cell culture media, wherein fibroblast cells of the skeletal muscle cell suspension adhere to the wall of the first container;
(b) re-plating non-adherent cells from step (a) in a second container containing media;
(c) re-plating the non-adherent cells from step (b) serially in 4 containers containing media, after 30-40% of cells have adhered to each container; and
(d) isolating the skeletal muscle-derived myoblasts from the wall of the sixth container following the re-plating in step (c), wherein the isolated cells are CD34 + , Bcl-2 + , KDR + , and Sca-1 + .
2. The method according to claim 1 , wherein the composition is introduced into the recipient by an administration route selected from injection or intravenous delivery.
3. The method according to claim 1 , wherein the composition further comprises a carrier, excipient, or diluent.
4. The method according to claim 3 , wherein the carrier comprises an absorbent or adherent material.
5. The method according to claim 3 , wherein the carrier is a collagen sponge material.
6. The method according to claim 1 , wherein the skeletal muscle-derived myoblasts are histocompatibly-matched with the recipient.
7. A method of treating fecal incontinence in a subject in need thereof, comprising introducing an isolated population of cultured cells enriched in viable, desmin-expressing, skeletal muscle-derived myoblasts into the anal sphincter soft-tissue of the recipient in an amount sufficient for the myoblasts to augment or bulk the anal sphincter soft-tissue; wherein the skeletal muscle-derived myoblasts are introduced in a composition comprising a physiologically acceptable medium, wherein the skeletal muscle-derived myoblasts are isolated using a method comprising:
(a) plating a suspension of skeletal muscle cells from skeletal muscle tissue in a first container containing cell culture media, wherein fibroblast cells of the skeletal muscle cell suspension adhere to the wall of the first container;
(b) re-plating non-adherent cells from step (a) in a second container containing media;
(c) re-plating the non-adherent cells from step (b) serially in 4 containers containing media, after 30-40% of cells have adhered to each container; and
(d) isolating the skeletal muscle-derived myoblasts from the wall of the sixth container following the re-plating in step (c), wherein the isolated cells are CD34 + , Bcl-2 + , KDR + , and Sca-1 + .
8. The method according to claim 7 , wherein the composition is introduced into the recipient by an administration route selected from injection or intravenous delivery.
9. The method according to claim 8 , wherein the composition further comprises a carrier, excipient, or diluent.
10. The method according to claim 9 , wherein the carrier comprises an absorbent or adherent material.
11. The method according to claim 10 , wherein the carrier is a collagen sponge material.
12. The method according to claim 8 , wherein the skeletal muscle-derived myoblasts are histocompatibly-matched with the recipient.