Method

Abstract

A method of preparing a therapeutic cell population for clinical use from a starting population of cells comprising haematopoietic stem cells, said method comprising separating a population of cells that substantially do not express CD38 but which express CD34 from the starting population of cells, and transducing the separated cell population with a vector to obtain the therapeutic cell population.

1 . A method of preparing a therapeutic cell population for clinical use from a starting population of cells comprising haematopoietic stem cells, said method comprising separating a population of cells that substantially do not express CD38 but which express CD34 from the starting population of cells, and transducing the separated cell population with a vector to obtain the therapeutic cell population. 
     
     
         2 . The method of  claim 1 , wherein:
 (i) the method comprises the steps of:
 (a) separating CD38-expressing cells from a starting population of cells comprising haematopoietic stem cells; 
 (b) separating CD34-expressing cells from the population of cells obtained in step (a) that do not express CD38; and 
 (c) transducing the CD34-expressing cell population obtained in step (b) with a vector to obtain the therapeutic cell population; or 
   (ii) the method comprises the steps of:
 (a) separating CD34-expressing cells from a starting population of cells comprising haematopoietic stem cells; 
 (b) separating CD38-expressing cells from the population of cells obtained in step (a) that express CD34; and 
 (c) transducing the cell population that does not express CD38 obtained in step (b) with a vector to obtain the therapeutic cell population. 
   
     
     
         3 . The method of  claim 1 , wherein the therapeutic cell population has the CD34 + CD38 −  phenotype. 
     
     
         4 . The method of  claim 1 , wherein the starting population of cells comprising haematopoietic stem cells is obtained from mobilised peripheral blood, bone marrow or umbilical cord blood. 
     
     
         5 . The method of  claim 1 , wherein the CD38-expressing cells and/or CD34-expressing cells are separated using magnetic bead-based separation or flow cytometry. 
     
     
         6 . The method of  claim 1 , wherein the vector is a viral vector, optionally wherein the viral vector is a lentiviral vector and/or comprises a nucleotide of interest. 
     
     
         7 . The method of  claim 1 , wherein the step of transducing a population of cells with a vector comprises:
 (a) culturing the cells for about 44 h or more, optionally for about 44-66 h, 44-60 h, 44-54 h or 44-48 h; or   (b) culturing the cells for less than about 44 h, optionally for about 12-42 h, 12-36 h, 12-24 h or 12-18 h.   
     
     
         8 . The method of  claim 1 , wherein the step of transducing a population of cells with a vector comprises contacting the population of cells with prostaglandin E2 or a prostaglandin E2 derivative, optionally wherein the cells are contacted with the prostaglandin E2 or prostaglandin E2 derivative before or at the same time as exposure of the cells to the vector. 
     
     
       9. The method of  claim 1 , wherein the separated CD38-expressing cells or portion thereof are retained to form a support cell population, optionally wherein the support cell population has the CD34 + CD38 int1 , CD34 + CD38 int2  and/or CD34 + CD38 +  phenotype. 
     
     
         10 . A method of treatment comprising administering a therapeutic cell population to a subject in need thereof, wherein the therapeutic cell population is prepared according to a method of preparing a therapeutic cell population for clinical use from a starting population of cells comprising haematopoietic stem cells, said method comprising separating a population of cells that substantially do not express CD38 but which express CD34 from the starting population of cells, and transducing the separated cell population with a vector to obtain the therapeutic cell population. 
     
     
         11 . The method of  claim 10 , wherein the therapeutic cell population is administered to a subject that has not undergone myeloablative conditioning, optionally wherein a support cell population is not administered to the subject. 
     
     
         12 . The method of  claim 10 , wherein the therapeutic cell population is administered in combination with a support cell population, wherein the support cell population is formed by retaining the separated CD38-expressing cells or portion thereof, optionally wherein the support cell population has the CD34 + CD38 int1 , CD34 + CD38 int2  and/or CD34 + CD38 +  phenotype, optionally wherein the cell populations are administered to a subject that has undergone myeloablative conditioning. 
     
     
         13 . The method of  claim 10 , wherein the therapeutic cell population is administered as part of an autologous stem cell transplant procedure or an allogeneic stem cell transplant procedure. 
     
     
         14 . The method of  claim 10 , wherein the therapeutic cell population is administered in combination with an allogeneic support cell population, optionally wherein the therapeutic cell population is an autologous or allogeneic cell population. 
     
     
         15 . The method of  claim 12 , wherein the therapeutic cell population is administered to a subject prior to administration of the support cell population, or wherein the therapeutic cell population is administered to a subject contemporaneously with or simultaneously to administration of the support cell population. 
     
     
         16 . The method of  claim 14 , wherein the therapeutic cell population is administered to a subject prior to administration of the support cell population, or wherein the therapeutic cell population is administered to a subject contemporaneously with or simultaneously to administration of the support cell population. 
     
     
         17 . The method of  claim 10 , wherein the therapeutic cell population has the CD34 + CD38 −  phenotype. 
     
     
         18 . The method of  claim 10 , wherein the step of transducing a population of cells with a vector comprises:
 (a) culturing the cells for about 44 h or more, optionally for about 44-66 h, 44-60 h, 44-54 h or 44-48 h; or   (b) culturing the cells for less than about 44 h, optionally for about 12-42 h, 12-36 h, 12-24 h or 12-18 h.   
     
     
         19 . The method of  claim 10 , wherein the step of transducing a population of cells with a vector comprises contacting the population of cells with prostaglandin E2 or a prostaglandin E2 derivative, optionally wherein the cells are contacted with the prostaglandin E2 or prostaglandin E2 derivative before or at the same time as exposure of the cells to the vector.