Isolated stromal cells and methods of using the same

Abstract

Methods of treating patients who are suffering from a disease, disorder or condition characterized by a bone cartilage or lung defect are disclosed. The methods comprising the step of intravenous administration of stromal cells isolated from normal syngeneic individuals or intravenous administration of stromal cells isolated from the patient subsequent to correction of the genetic defect in the isolated cells. Implant devices comprising a container that has at least one membrane surface and stromal cells isolated from bone marrow that comprise a gene construct are disclosed. The gene construct in the stromal cells comprises a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in stromal cells. Methods of treating individuals with diseases, disorders or conditions which can be treated with a beneficial protein, including diseases, disorders or conditions characterized by gene defects are disclosed. The methods comprise introducing into such individuals, stromal cells that are administered in a manner that physically isolates them from the recipient's immune system and that comprise a gene construct that comprises a nucleotide sequence that encodes a beneficial protein operably linked to regulatory elements which function in stromal cells.

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 .- 11 . (canceled) 
   
   
       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 .- 21 . (canceled) 
   
   
       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 .- 24 . (canceled) 
   
   
       25 . The implant device of  claim 22  comprising 10 4  to 10 11  stromal cells. 
   
   
       26 . (canceled) 
   
   
       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 .- 30 . (canceled) 
   
   
       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 .- 36 . (canceled) 
   
   
       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 . (canceled) 
   
   
       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 .- 44 . (canceled) 
   
   
       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 .- 54 . (canceled)