CPC A61K 35/15 (2013.01) [A61K 38/177 (2013.01); C07K 16/00 (2013.01); C07K 16/30 (2013.01); C07K 16/32 (2013.01); C12N 5/0645 (2013.01); C07K 2317/622 (2013.01); C07K 2319/03 (2013.01); C07K 2319/33 (2013.01); C12N 2501/599 (2013.01); C12N 2510/00 (2013.01)] | 25 Claims |
1. A method of making a population of chimeric antigen receptor (CAR) encoded CD14+ cells, the method comprising:
(a) extracting a leukapheresis sample or a PBMC sample from a human subject;
(b) isolating CD14+ cells from CD3+ cells in the leukapheresis sample from (a) or the PBMC sample from (a);
(c) after (b), culturing the isolated CD14+ cells ex vivo in the presence of (i) a growth factor, wherein the growth factor comprises recombinant human M-CSF and does not comprise GM-CSF and (ii) a cytokine that does not comprise IL-2,
thereby obtaining an ex vivo population of CD14+ cells; and
(d) introducing a recombinant polynucleic acid encoding a chimeric antigen receptor (CAR) into the ex vivo population of CD14+ cells, thereby obtaining the population of CAR encoded CD14+ cells;
wherein the CAR comprises (i) an extracellular domain comprising an antigen binding domain; (ii) a transmembrane domain; and (iii) an intracellular domain comprising a CD3 zeta intracellular signaling domain.
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