END-TO-END CELL THERAPY AUTOMATION
The present disclosure provides an automated method of producing genetically modified immune cells, including chimeric antigen receptor T (CAR T) cells, utilizing a fully-enclosed cell engineering system.
1 . A cassette for use in an automated cell engineering system, the cassette comprising:
a low temperature chamber for storage of a cell culture media;
a high temperature chamber for carrying out activation, transduction and expansion of an immune cell culture; and
a thermal barrier, wherein the high temperature chamber is separated from the low temperature chamber by the thermal barrier,
wherein the high temperature chamber includes a cell culture chamber and one or more fluidics pathways connected to the cell culture chamber, and
wherein the fluidics pathways provide recirculation, removal of waste and homogenous gas exchange and distribution of nutrients to the cell culture chamber without disturbing cells within the cell culture chamber.
2 . The cassette of claim 1 , wherein the low temperature chamber is maintained from about 4° C. to about 8° C.
3 . The cassette of claim 1 , wherein the high temperature chamber is maintained from about 35° C. to about 40° C.
4 . The cassette of claim 1 , wherein the low temperature chamber can further include a bag or other holder for media, including about 1 L or more of fluid.
5 . The cassette of claim 1 , wherein the cassette further includes one or more pumps for driving fluid through the cassette.
6 . The cassette of claim 1 , wherein the cassette further includes one or more valves for controlling the flow through the fluidic pathways.
7 . The cassette of claim 1 , wherein the cassette further includes one or more sampling ports and/or injection ports.
8 . The cassette of claim 1 , wherein the cassette further includes one or more of a pH sensor, a glucose sensor, an oxygen sensor, a carbon dioxide sensor, a lactic acid sensor and/or an optical density sensor.
9 . A cassette for use in an automated cell engineering system, the cassette comprising:
a low temperature chamber for storage of a cell culture media; and
a high temperature chamber for carrying out activation, transduction and expansion of an immune cell culture,
wherein the high temperature chamber includes:
a flat and non-flexible cell culture chamber having a chamber height of less than 5.0 cm, and
one or more fluidics pathways connected to the cell culture chamber, wherein the fluidics pathways provide recirculation, removal of waste and homogenous gas exchange and distribution of nutrients to the cell culture chamber without disturbing cells within the cell culture chamber.
10 . The cassette of claim 9 , wherein the chamber height is about 0.5 cm to about 4.0 CM.
11 . The cassette of claim 9 , wherein the cell culture chamber has a volume of about 0.5 mL to about 300 mL.
12 . The cassette of claim 9 , wherein the cell culture chamber further comprises at least one of:
a distal port configured to allow for the removal of air bubbles from the cell culture chamber and/or as a recirculation port;
a medial port configured to function as a recirculation inlet port; and
a proximal port configured to function as a drain port for cell removal.
13 . The cassette of claim 9 , wherein the cassette further includes one or more of a pH sensor, a glucose sensor, an oxygen sensor, a carbon dioxide sensor, a lactic acid sensor and/or an optical density sensor.
14 . A cassette for use in an automated cell engineering system, the cassette comprising:
a low temperature chamber for storage of a cell culture media; and
a high temperature chamber for carrying out activation, transduction and expansion of an immune cell culture,
wherein the high temperature chamber includes:
a cell culture chamber; and
one or more fluidics pathways connected to the cell culture chamber, wherein the fluidics pathways provide recirculation, removal of waste and homogenous gas exchange and distribution of nutrients to the cell culture chamber without disturbing cells within the cell culture chamber, and
wherein the fluidics pathways comprise a silicon-based tubing component that allows oxygenation through the tubing component.
15 . The cassette of claim 14 , wherein the low temperature chamber can further include a bag or other holder for media, including about 1 L or more of fluid.
16 . The cassette of claim 14 , wherein the cassette further includes one or more pumps for driving fluid through the cassette.
17 . The cassette of claim 14 , wherein the cassette further includes one or more valves for controlling the flow through the fluidic pathways.
18 . The cassette of claim 14 , wherein the cassette further includes one or more sampling ports and/or injection ports.
19 . The cassette of claim 14 , wherein the cassette further includes one or more of a pH sensor, a glucose sensor, an oxygen sensor, a carbon dioxide sensor, a lactic acid sensor and/or an optical density sensor.