Patent ID: 8685385

Claim:
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 + .