US 12,029,763 B2
Immuno-modulatory progenitor (IMP) cell
Ajan Reginald, Swansea (GB); Martin John Evans, Swansea (GB); and Sabena Sultan, Swansea (GB)
Assigned to CELL THERAPY LIMITED, (GB)
Appl. No. 14/904,411
Filed by CELL THERAPY LIMITED, Swansea (GB)
PCT Filed Jun. 9, 2015, PCT No. PCT/GB2015/051673
§ 371(c)(1), (2) Date Jan. 11, 2016,
PCT Pub. No. WO2015/189587, PCT Pub. Date Dec. 17, 2015.
Claims priority of application No. 1410504 (GB), filed on Jun. 12, 2014.
Prior Publication US 2017/0080029 A1, Mar. 23, 2017
Int. Cl. A61K 35/28 (2015.01); C12N 5/071 (2010.01); C12N 5/0775 (2010.01); C12N 5/078 (2010.01)
CPC A61K 35/28 (2013.01) [C12N 5/0633 (2013.01); C12N 5/0634 (2013.01); C12N 5/0663 (2013.01); C12N 5/0665 (2013.01); C12N 2500/02 (2013.01); C12N 2500/84 (2013.01); C12N 2500/90 (2013.01); C12N 2506/1353 (2013.01)] 3 Claims
 
1. A method of producing a population of IMP cells, comprising
(a) physically separating mononuclear cells (MCs) from bone marrow and culturing the MCs for 15 to 25 days in a medium consisting essentially of alpha Minimum Essential Medium (MEM), L-alanyl-L-glutamine dipeptide, platelet lysate, and heparin, at 0% oxygen (O2) supplied by an incubator and under conditions which allow the MCs to adhere to induce the MCs to differentiate into IMP cells and
(b) harvesting and culturing the IMP cells in (a) to thereby produce the population, wherein harvesting comprises an antibody-based technique to select for a population in which
(i) at least 90% of the cells in the population express detectable levels of MIC A/B,
(ii) at least 60% of the cells in the population express detectable levels of CD304 (Neuropilin 1),
(iii) at least 45% of the cells in the population express detectable levels of CD178 (FAS ligand),
(iv) at least 10% of the cells in the population express detectable levels of CD289 (Toll-like receptor 9),
(v) at least 15% of the population express detectable levels of CD363 (Sphingosine-1-phosphate receptor 1),
(vi) at least 20% of the cells in the population express detectable levels of CD99,
(vii) at least 80% of the cells in the population express detectable levels of CD181 (C-X-C chemokine receptor type 1; CXCR1),
(viii) at least 30% of the cells in the population express detectable levels of epidermal growth factor receptor (EGF-R),
(ix) at least 60% of the cells in the population express detectable levels of CXCR2
(x) at least 5% of the cells in the population express detectable levels of CD126,
(xi) at least 90% of the cells in the population express detectable levels of HLA-ABC,
(xii) at least 90% of the cells in the population express detectable levels of CD340,
(xiii) the cells express detectable levels of CD83,
(xiv) the cells express detectable levels of CD86, and
(xv) 1% or fewer of the cells express detectable levels of CD70 and wherein the platelet lysate is produced by subjecting a population of human platelets to four freeze-thaw cycles, wherein the freeze portion of each cycle is carried out in liquid nitrogen.