Process for obtaining functional lymphocytes cells

A method for differentiating an iPSc into a functional TEP, the method comprising the following steps:

• a) incubating the iPSc in a culture medium supplemented with a (R)-(+)-trans-4-(1-Aminoethyl)-N-(4-Pyridyl)cyclohexanecarboxamide dihydrochloride (Y27) compound for 24 h, to obtain a Y27 culture medium,
• b) removing the Y27 culture medium from the cell, and incubating for 24 h the cell with a second culture medium supplemented with an Activin A growth factor and a -[[2-[[4-(2,4-Dichlorophenyl)-5-(5-methyl-1H-imidazol-2-yl)-2-pyrimidinyl]amino]ethyl]amino]-3-pyridinecarbonitrile (CHIR99) compound, to obtain the cell in an ActA/CHIR99 culture medium,
• c) removing the ActA/CHIR99 culture medium from the cell, and incubating for 48 h the cell with a third culture medium supplemented with an Actinin A growth factor to obtain the cell in an ActA culture medium,
• d) removing the ActA culture medium from the cell, and incubating for 24 h the cell in a fourth medium supplemented with an Actinin A growth factor and a Y27 compound, to obtain the cell in a Y27/ActA culture medium,
• e) removing the Y27/ActA culture medium from the cell, and incubating for 24 h the cell in a fifth medium supplemented with a fibroblast growth factor 8 (FGF8) growth factor and a retinoic acid (RA) compound, to obtain the cell in a RA/F8 culture medium,
• f) removing the RA/F8 culture medium from the cell, and incubating for 48 h the cell in a sixth medium supplemented with a FGF8 growth factor, a retinoic acid compound, a Noggin growth factor and a 4-[3-(2-pyridinyl)-1H-pyrazol-4-yl]-quinoline (LY3) compound for 48 h, to obtain the cell in a RA/F8/NOG/LY3 culture medium,
• g) removing the RA/F8/NOG/LY3 culture medium from the cell, and incubating for 48 h the cell in a seventh medium supplemented with a FGF8 growth factor, a retinoic acid compound, a bone morphogenetic protein 4 (BMP4) growth factor, a LY3 compound and a CHIR99 compound, to obtain the cell in a RA/F8/LY3/CHIR99/BMP culture medium,
• h) removing the RA/F8/LY3/CHIR99/BMP culture medium from the cell, and incubating for 24 h the cell in an eighth medium supplemented with a FGF8 growth factor, a retinoic acid compound, a BMP4 growth factor and a CHIR99 compound, to obtain the cell in a first RA/F8/CHIR99/BMP culture medium,
• i) removing the first RA/F8/CHIR99/BMP culture medium from the cell, and incubating for 24 h the cell in a nineth medium supplemented with a FGF8 growth factor, a retinoic acid compound, a bone morphogenetic protein 4 (BMP4) growth factor and a CHIR99 compound to obtain the cell in a second RA/F8/CHIR99/BMP culture medium,
• j) removing the second RA/F8/CHIR99/BMP culture medium from the cell, and incubating for 72 h the cell in a tenth medium supplemented with a FGF8 growth factor, a BMP4 growth factor, a fibroblast growth factor 10 (FGF 10) growth factor, an Insulin-like growth factor 1 (IGF1) growth factor and an Epidermal growth factor (EGF) growth factor, to obtain the cells in a RA/F8/BMP/F10/IGF1/EGF culture medium, and
• k) recovering the functional TEP from the RA/F8/BMP/F10/IGF1/EGF culture medium, after 72 h.