Cardiac neural crest cells and methods of use thereof

Abstract

Cardiac neural crest cells and methods for making and using the same to effect cardiac injury is described herein.

1 . A method for treating a cardiac injury in a subject in need thereof comprising administering a therapeutically effective amount of a pharmaceutical composition comprising cardiac neural crest cells. 
     
     
         2 . The method of  claim 1 , wherein the cardiac neural crest cells are cKit positive. 
     
     
         3 . The method of  claim 1 , wherein the cardiac injury is a cardiac injury caused by myocardial infarction, heart failure, cardiomyopathy, congenital heart disease, nutritional diseases, ischemic or non-ischemic cardiomyopathy, hypertensive cardiomyopathy, valvular cardiomyopathy, inflammatory cardiomyopathy, cardiomyopathy secondary to a systemic metabolic disease, alcoholic cardiomyopathy, diabetic cardiomyopathy, restrictive cardiomyopathy, and combinations thereof. 
     
     
         4 . The method of  claim 1 , wherein the pharmaceutical composition has about 50% by weight to about 100% by weight cardiac neural crest cells. 
     
     
         5 . The method of  claim 1 , wherein the pharmaceutical composition has about 50% by weight cardiac neural crest cells. 
     
     
         6 - 7 . (canceled) 
     
     
         8 . The method of  claim 1 , wherein the pharmaceutical composition has about 1×10 6  cardiac neural crest cells. 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 1 , wherein the pharmaceutical composition further comprises one or more differentiation agents. 
     
     
         11 . The method of  claim 10 , wherein the one or more differentiation agents are selected from BMP antagonist, noggin, chordin, Tsg, soluble receptors such as BMPR1A and BMPR1B, small molecule BMP antagonist such as dorsomorphin, retinoic acid receptor agonist such as retinoic acid, vitamin A, LG100268, or LGD1069, wnt inhibitor such as dickkopf homolog 1 (DKK1). 
     
     
         12 . The method of  claim 1 , wherein the cardiac neural crest cells have a concentration of stem cell factor-receptors of about 50 receptors/micrometer 2 to about 800 receptors/micrometer 2 . 
     
     
         13 . The method of  claim 1 , wherein the cardiac neural crest cells have a concentration of stem cell factor-receptors of about 200 receptors/micrometer 2  to about 600 receptors/micrometer 2 . 
     
     
         14 . A method of producing cardiac neural crest cells comprising:
 culturing a plurality of stem cells with noggin and leukemia inhibitory factor;   culturing the plurality of stem cells with noggin, thereby forming three-dimensional aggregates of the plurality of stem cells; and   culturing the plurality of stem cells in cardiac neural crest differentiation medium to form cardiac neural crest cells.   
     
     
         15 . The method of  claim 14 , wherein the cardiac neural crest cells express stem cell factor-receptor. 
     
     
         16 . The method of  claim 14 , wherein the cardiac neural crest cells are cKit positive. 
     
     
         17 . The method of  claim 14 , wherein the plurality of stem cells are induced pluripotent stem cells. 
     
     
         18 . The method of  claim 14 , wherein the plurality of stem cells are embryonic stem cells. 
     
     
         19 . The method of  claim 14 , wherein the noggin in the step of culturing a plurality of stem cells with noggin and leukemia inhibitory factor is at a concentration of 150 ng/ml. 
     
     
         20 . The method of  claim 14 , wherein the leukemia inhibitory factor in the step of culturing a plurality of stem cells with noggin and leukemia inhibitory factor is at a concentration of 2000 units/ml. 
     
     
         21 . The method of  claim 14 , wherein the cardiac neural crest cells have a concentration of stem cell factor-receptors of about 50 receptors/micrometer 2  to about 800 receptors/micrometer 2 . 
     
     
         22 . (canceled) 
     
     
         23 . The method of  claim 14 , further comprising contacting tamoxifen to the stem cells. 
     
     
         24 . A pharmaceutical composition comprising cardiac neural crest cells and a pharmaceutically acceptable carrier or excipient. 
     
     
         25 . (canceled) 
     
     
         26 . The pharmaceutical composition of  claim 24 , wherein the cardiac neural crest cells have a concentration of stem cell factor-receptors of about 50 receptors/micrometer 2  to about 800 receptors/micrometer 2 . 
     
     
         27 . (canceled)