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 improving function of bladder of a subject in need thereof, comprising:
introducing an enriched population of autologous, skeletal muscle-derived progenitor cells (MDCs) into a wall of a bladder muscle tissue of a recipient, in an amount effective to improve bladder function; wherein
the MDCs are isolated by a method comprising:
(i) plating a suspension of cells from skeletal muscle tissue in a first container containing cell culture media, wherein fibroblast cells in the muscle tissue suspension adhere to the wall of the first container;
(ii) after about one hour, re-plating non-adherent cells from (i) in a fresh second container containing cell culture media similar to the first container, wherein 30-40% of the non-adherent cells transferred from the first container adhere to the wall of the second container;
(iii) re-plating the non-adherent cells from step (ii) at least one time, after 30-40% of cells have adhered to the wall of the second or the subsequent containers, to enrich for an end population of viable, non-fibroblast, desmin-expressing, skeletal muscle-derived progenitor cells (MDCs) adhered to the wall of the containers; and
(iv) isolating the MDCs from the wall of the containers, wherein the MDCs are desmin and CD34 positive.
2. The method according to claim 1 , wherein the MDCs are introduced in a composition comprising a physiologically acceptable medium.
3. 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 bladder wall to be treated in a physiologically acceptable medium.
4. The method according to claim 1 , wherein a cloned population of the MDCs is introduced into the recipient.
5. The method according to claim 1 , wherein the MDCs are contacted with a cytokine or growth factor chosen from basic fibroblast growth factor (b-FGF), insulin-like growth factor (IGF), or nerve growth factor (NGF), prior to introducing the skeletal muscle-derived myoblasts into the recipient.
6. A method of improving contractility of bladder smooth muscle, comprising:
introducing an enriched population of autologous, skeletal muscle-derived progenitor cells (MDCs) into the bladder wall smooth muscle of a recipient in an amount effective to improve contractility of bladder smooth muscle; wherein the MDCs are isolated by a method comprising:
(i) plating a suspension of cells from skeletal muscle tissue in a first container containing cell culture media, wherein fibroblast cells in the muscle tissue suspension adhere to the wall of the first container;
(ii) after about one hour, re-plating non-adherent cells from (i) in a fresh second container containing cell culture media similar to the first container, wherein 30-40% of the non-adherent cells transferred from the first container adhere to the wall of the second container;
(iii) re-plating the non-adherent cells from step (ii) at least one time, after 30-40% of cells have adhered to the wall of the second or the subsequent containers, to enrich for an end population of viable, non-fibroblast, desmin-expressing, skeletal muscle-derived progenitor cells (MDCs) adhered to the wall of the containers; and
(iv) isolating the MDCs from the wall of the containers, wherein the MDCs are desmin and CD34 positive.
7. The method according to claim 6 , wherein the MDCs are introduced in a composition comprising a physiologically acceptable medium.
8. The method according to claim 6 , wherein the MDCs are introduced in an amount of about 10 5 to 10 6 cells per cm 3 of bladder wall smooth muscle to be treated in a physiologically acceptable medium.
9. The method according to claim 6 , wherein a cloned population of the MDCs is introduced into the recipient.
10. The method according to claim 6 , wherein the MDCs are contacted with a cytokine or growth factor selected from one or more of basic fibroblast growth factor (b-FGF), insulin-like growth factor (IGF), or nerve growth factor (NGF), prior to introducing the skeletal muscle-derived myoblasts into the recipient.