| CPC C12M 23/04 (2013.01) [C12M 23/20 (2013.01); C12M 23/24 (2013.01); C12M 23/42 (2013.01); C12M 27/00 (2013.01); C12M 29/24 (2013.01); C12M 41/12 (2013.01); C12M 41/32 (2013.01); C12M 41/34 (2013.01); C12M 41/48 (2013.01); C12N 5/0068 (2013.01); G01N 35/1009 (2013.01); C12N 2533/52 (2013.01)] | 20 Claims |

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1. A cassette for use in an automated cell engineering system as a removable and replaceable element, comprising:
a. a high temperature chamber configured for carrying out activation, transduction and/or expansion of a cell culture at a temperature of between 35° C. to 40° C., the high temperature chamber including a cell culture chamber;
b. a low temperature chamber configured for storage of a reagent at a temperature of between 4° C. to 8° C.;
c. a thermal barrier adjacent the high temperature chamber and the low temperature chamber and separating the high temperature chamber from the low temperature chamber within the cassette; and
d. 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;
wherein the cell culture chamber is a flat and non-flexible chamber, having a low chamber height and a cell-contacting surface, and the cell culture chamber is maintained in a substantially planar orientation in the cassette, and
wherein at least a portion of the cell-contacting surface comprises a non-porous, gas-permeable material, and the cell-contacting surface further includes multiple structures cut or made into the cell-contacting surface to add surface area to increase cell adhesion, wherein the multiple structures are sandwiched between sheets of the non-porous, gas-permeable material.
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