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 restoring or improving contractility of a smooth muscle tissue in a mammal, comprising
(a) isolating the skeletal muscle-derived progenitor cells (MDCs) according to a culture method comprising:
(i) plating a suspension of cells from skeletal muscle tissue in a cell culture media in a first container, to which fibroblast cells in the muscle tissue suspension adhere;
(ii) re-plating non-adherent cells in the cell culture media from step (i) in a second container containing media after approximately 15% to 20% of the cells from the cell suspension have adhered to the first container;
(iii) re-plating the non-adherent cells in step (ii) at least one time in another cell culture container to enrich for an end population of viable, non-fibroblast, desmin-expressing said MDCs; and
(iv) isolating said end population of viable, non-fibroblast, desmin-expressing MDCs from the container in step (iii); and
(b) administering to said smooth muscle tissue a physiologically acceptable composition comprising isolated MDCs in an amount sufficient to restore or improve contractility of said smooth muscle, and a carrier, excipient or diluents, wherein the physiologically acceptable composition bulks, fills, or supports the smooth muscle tissue.
2. The method according to claim 1 , wherein the MDCs express CD34.
3. The method according to claim 1 , wherein the MDCs express Bcl-2.
4. The method according to claim 1 , wherein the MDCs express CD34 and Bcl-2.
5. The method according to claim 1 , wherein the MDCs are a clonal population of muscle-derived progenitor cells.
6. The method according to claim 1 , wherein the MDCs are histocompatibility-matched with the recipient.
7. The method according to claim 1 , wherein the MDCs are introduced in an amount of about 10 5 to 10 6 cells per cm 3 of the smooth muscle tissue.