Isolated stromal cells and methods of using the same

Abstract

Isolated stromal cells, containers comprising isolated stromal cells transfected with exogenous DNA, and methods of treating patients suffering from diseases characterized by a bone cartilage or lung defect are disclosed. The methods comprise obtaining a bone marrow sample from a donor, isolating stromal cells from the sample, and administering the isolated stromal cells to the patient.

1 . A method of treating patient who is suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect comprising the steps of:
 a) obtaining a bone marrow sample from a donor who is not suffering from a disease, disorder or condition characterized by a bone, cartilage or lung defect and who is syngeneic with said patient;   b) isolating stromal cells from said sample; and,   c) administering said isolated stromal cells by intravenous infusion to said patient.   
     
     
         2 . The method of  claim 1  wherein said patent undergoes bone marrow ablation prior to administration of isolated stromal cells. 
     
     
         3 . The method of  claim 2  wherein said stromal cells are administered by intravenous infusion to said patient together with hematopoeitic precursor cells from a bone marrow sample from a donor who is not suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect and who is syngeneic with said patient. 
     
     
         4 . The method of  claim 2  wherein said stromal cells are administered by intravenous infusion to said patient free from hematopoeitic precursor cells. 
     
     
         5 . The method of  claim 1  wherein prior to administering said stromal cells, said stromal cells are transfected with a gene construct that comprises a herpes thymidine kinase gene, wherein said gene is operably linked to regulatory sequences and is expressed by said stromal cells. 
     
     
         6 . The method of  claim 1  wherein said disease, disorder or condition is characterized by a defect in said patient's bone. 
     
     
         7 . The method of  claim 6  wherein said disease, disorder or condition is osteogenesis imperfecta or osteoporosis. 
     
     
         8 . The method of  claim 1  wherein said disease, disorder or condition is characterized by a defect in said patient's cartilage. 
     
     
         9 . The method of  claim 8  wherein said disease, disorder or condition is chondrodysplasia or osteoarthritis. 
     
     
         10 . The method of  claim 1  wherein said disease, disorder or condition is characterized by defect in said patient's lungs. 
     
     
         11 . The method of  claim 10  wherein said disease, disorder or condition characterized is cystic fibrosis. 
     
     
         12 . A method of treating patient who suffering from a disease, disorder or condition characterized by a mutated, non-functioning or under-expressed gene which results in a defect in the bone, cartilage or lungs of said patient comprising the steps of:
 a) obtaining a bone marrow sample from said patient;   b) isolating stromal cells from said sample;   c) transfecting said stromal cells with a normal copy of said mutated, non-functioning or under-expressed gene wherein said copy of said gene is operably linked to functional regulatory elements; and   d) administering said transfected stromal cells to said patient by intravenous infusion.   
     
     
         13 . The method of  claim 12  wherein said patent undergoes bone marrow ablation prior to administration of stromal cells. 
     
     
         14 . The method of  claim 13  wherein said stromal cells are administered by intravenous infusion to said patient together with hematopoietic precursor cells from said sample. 
     
     
         15 . The method of  claim 12  wherein prior to administering said stromal cells, said stromal cells are transfected with a gene construct that comprises a herpes thymidine kinase gene, wherein said gene is operably linked to regulatory sequences and is expressed by said stromal cells. 
     
     
         16 . The method of  claim 12  wherein said disease, disorder or condition is characterized by a defect in said patient's bone. 
     
     
         17 . The method of  claim 16  wherein said disease, disorder or condition is osteogenesis imperfecta and said gene encodes type I procollagen or type I collagen. 
     
     
         18 . The method of  claim 12  wherein said disease, disorder or condition is characterized by a defect in said patient's cartilage. 
     
     
         19 . The method of  claim 18  wherein said disease, disorder or condition is chondrodysplasia and said gene encodes type II procollagen or type II collagen. 
     
     
         20 . The method of  claim 12  wherein said disease, disorder or condition is characterized by defect in said patient's lungs. 
     
     
         21 . The method of  claim 20  wherein said disease, disorder or condition characterized is cystic fibrosis and said gene is a cystic fibrosis gene. 
     
     
         22 . An implant device comprising:
 a container having at least one membrane surface   stromal cells that comprise a gene construct, said gene construct comprising a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in said stromal cell.   
     
     
         23 . The implant device of  claim 22  wherein said membrane has a pore size of 0.3 microns. 
     
     
         24 . The implant device of  claim 22  having a membrane surface area of at least 100 mm 2 . 
     
     
         25 . The implant device of  claim 22  comprising 10 4  to 10 11  stromal cells. 
     
     
         26 . The implant device of  claim 22  comprising 10 4  to 10 8  stromal cells. 
     
     
         27 . The implant device of  claim 22  wherein said beneficial protein is selected form the group consisting of human growth hormone, obesity factor and human Factor VIII. 
     
     
         28 . A method of treating an individual with a disease, disorder or condition which can be treated with a beneficial protein comprising the step of introducing into such an individual, immunologically isolated stromal cells that comprise a gene construct, said gene construct comprising a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in said stromal cell. 
     
     
         29 . The method of  claim 28  wherein said disease, disorder or condition which can be treated with a beneficial protein is a disease, disorder or conditions characterized by a gene defect. 
     
     
         30 . The method of  claim 29  wherein said beneficial protein is selected from the group consisting of human growth hormone and human Factor VIII. 
     
     
         31 . The method of  claim 28  wherein said immunologically isolated stromal cells are within an implant device that comprises said stromal cells and a container having at least one membrane surface. 
     
     
         32 . The method of  claim 31  wherein said membrane of said implant device has a pore size of 0.3 microns. 
     
     
         33 . The method of  claim 31  wherein said implant device has a membrane surface area of at least 100 mm 2 . 
     
     
         34 . The method of  claim 31  wherein said implant device comprises 10 4  to 10 11  stromal cells. 
     
     
         35 . The method of  claim 31  wherein said implant device comprises 10 4  to 10 8  stromal cells. 
     
     
         36 . The method of  claim 31  wherein said implant device is implanted into said individual subcutaneously. 
     
     
         37 . Immunologically isolated stromal cells that comprise a gene construct, said gene construct comprising a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in said stromal cell. 
     
     
         38 . The immunologically isolated stromal cells of  claim 37  wherein said stromal cells are microencapsulated. 
     
     
         39 . A method of treating patient who is suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect comprising the steps of:
 a) obtaining a bone marrow sample from a donor who is not suffering from a disease, disorder or condition characterized by a bone or cartilage defect and who is syngeneic with said patient; and,   b) administering a therapeutically effective amount of said bone marrow by intravenous infusion to said patient.   
     
     
         40 . The method of  claim 39  wherein said patent undergoes bone marrow ablation prior to administration of isolated stromal cells. 
     
     
         41 . The method of  claim 39  wherein said disease, disorder or condition is characterized by a defect in said patient's bone. 
     
     
         42 . The method of  claim 41  wherein said disease, disorder or condition is osteogenesis imperfecta. 
     
     
         43 . The method of  claim 39  wherein said disease, disorder or condition is characterized by a defect in said patient's cartilage. 
     
     
         44 . The method of  claim 43  wherein said disease, disorder or condition is chondrodysplasia. 
     
     
         45 . A method of treating patient who suffering from a disease, disorder or condition characterized by a mutated, non-functioning or under-expressed gene which results in a defect in the bone, cartilage or lungs of said patient comprising the steps of:
 a) obtaining a bone marrow sample from said patient;   b) isolating stromal cells from said sample;   c) culturing said stromal cells under conditions which result in replication of said stromal cells into an expanded culture of stromal cells; and   d) administering stromal cells of said expanded culture of stromal cells to said patient by intravenous infusion.   
     
     
         46 . The method of  claim 45  wherein said patent undergoes bone marrow ablation prior to administration of stromal cells. 
     
     
         47 . The method of  claim 46  wherein said stromal cells are administered by intravenous infusion to said patient together with hematopoietic precursor cells from said sample. 
     
     
         48 . The method of  claim 46  wherein said stromal cells are administered by intravenous infusion to said patient free from precursor cells from said sample. 
     
     
         49 . The method of  claim 45  wherein said disease, disorder or condition is characterized by a defect in said patient's bone. 
     
     
         50 . The method of  claim 49  wherein said disease, disorder or condition is osteogenesis imperfecta or osteoporosis. 
     
     
         51 . The method of  claim 45  wherein said disease, disorder or condition is characterized by a defect in said patient's cartilage. 
     
     
         52 . The method of  claim 46  wherein said disease, disorder or condition is chondrodysplasia or osteoarthritis. 
     
     
         53 . The method of  claim 45  wherein said disease, disorder or condition is characterized by defect in said patient's lungs. 
     
     
         54 . The method of  claim 53  wherein said disease, disorder or condition characterized is cystic fibrosis and said gene is a cystic fibrosis gene.