Method for identifying, expanding, and removing adult stem cells and cancer stem cells

Abstract

The invention relates to the fields of biochemistry, pharmacy and oncology. The invention particularly relates to the use of novel stem cell markers for the isolation of stem cells. The invention further relates to the obtained stem cells and their use in for example research or treatment, for example, for the preparation of a medicament for the treatment of damaged or diseased tissue. In one of the embodiments, the invention provides a method for obtaining (or isolating) stem cells comprising optionally preparing a cell suspension from a tissue or organ sample, contacting said cell suspension with an Lgr 6 or 5 binding compound, identify the cells bound to said binding compound, and optionally isolating the stem cells from said binding compound. The invention further relates to means suitable for cancer treatment and even more specific for the treatment of cancer by eradicating cancer stem cells.

1 - 58 . (canceled) 
     
     
         59 . An in vitro or ex vivo method of maintaining or culturing tissue or organ stem cells comprising providing the tissue or organ stem cells with an Lgr5 and/or Lgr6 binding compound. 
     
     
         60 . The method of  claim 59 , wherein binding compound is an antibody or an antibody derivative or an antibody fragment capable of binding to Lgr5 and/or Lgr6. 
     
     
         61 . The method of  claim 60 , wherein the antibody is a monoclonal antibody. 
     
     
         62 . The method of  claim 60 , wherein the antibody fragment is scFv, Fab, or (Fab)2 fragment. 
     
     
         63 . The method of  claim 60 , wherein the antibody derivative is a chimeric antibody, nanobody, bifunctional antibody, or humanized antibody. 
     
     
         64 . The method of  claim 59 , wherein the binding compound is an Lgr5 or Lgr6 ligand. 
     
     
         65 . The method of  claim 59 , wherein the binding compound is a small molecule agonist of Lgr5 or Lgr6. 
     
     
         66 . The method of  claim 59 , wherein the stem cells are adult stem cells. 
     
     
         67 . The method of  claim 59 , wherein the stem cells are isolated stem cells. 
     
     
         68 . The method of  claim 67 , further comprising isolating the stem cells from a cell suspension by:
 providing a cell suspension;   contacting said cell suspension with an antibody specific for Lgr5 so as to bind the antibody specific for Lgr5 to Lgr5 positive cells in the cell suspension; and   at least partially purifying cells bound to the antibody specific for Lgr5 from cells not bound to the antibody specific for Lgr5 from the cell suspension so as to obtain the stem cells.   
     
     
         69 . The method of  claim 59 , wherein the stem cells are intestine, brain, lung, heart, skin, liver, retina, stomach, pancreas, ovary, adrenal medulla, bladder, bone, connective tissue, ear, muscle, prostate, placenta, uterus, or breast stem cells. 
     
     
         70 . The method of  claim 59 , wherein the stem cells are cancer stem cells. 
     
     
         71 . The method of  claim 70 , wherein the cancer stem cells are obtained from a tumor and the method further comprises determining the cancer stem cell content of the tumor. 
     
     
         72 . The method of  claim 70 , wherein the cancer stem cells are comprised in a tumor and the method further comprises determining the cancer stem cell content of the tumor. 
     
     
         73 . The method of  claim 71 , wherein the cancer stem cells are comprised in a tumor and the method further comprises determining the cancer stem cell content of the tumor. 
     
     
         74 . The method of  claim 72 , wherein determining the cancer stem cell content of the tumor comprises contacting the tumor with the Lgr5 and/or Lgr6 binding compound or a second Lgr5 and/or Lgr6 binding compound, removing unbound Lgr5 and/or Lgr6 binding compound and determining whether any bound binding compound is present in the tumour. 
     
     
         75 . The method of  claim 73 , wherein determining the cancer stem cell content of the tumor comprises contacting the tumor with the Lgr5 and/or Lgr6 binding compound or a second Lgr5 and/or Lgr6 binding compound, removing unbound Lgr5 and/or Lgr6 binding compound and determining whether any bound binding compound is present in the tumour.