This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells. The present compositions and methods entail viral delivery of an editing cassette to live mammalian cells such that the editing cassettes edit the cells and the edited cells continue to grow, preferably using a fully-automated end-to-end instrument to process the cells without human intervention to enhance cell processing uniformity and to maintain the integrity of the cell culture.

A bioreactor includes a base and a lid. The base includes two opposing curved or convoluted surfaces, two opposing flat surfaces, a window disposed on one of the flat surfaces, the window having a higher degree of transparency compared to other portions of the base, wherein images or videos of cells are captured through the window by non-invasive ISM device, and a rounded bottom. The lid is attachable to the base. The lid includes a shaft and a semipermeable membrane attached to the shaft, the semipermeable membrane being permeable to oxygen but impermeable to viruses and bacteria. The PAT-based online analysis directs the adaptive manipulations of bioreactor towards efficient, automated, and GMP-compliant clinical protocols of cell culture.

Cell culture apparatus for emulating gastrointestinal tract conditions and comprising at least two adjacent, microfluidic, cell cultivation channels separated by a permeable or semipermeable membrane, a first channel carrying gastrointestinal tract epithelial cells or tissues and a second channel carrying luminal and preferably mucosal microbiota, and wherein said second channel comprises one or more dwell chambers capable of providing a location for unattached luminal flora to reside away from any direct flow in said second channel, permits modelling of multiple sections of the gastrointestinal tract and control of retention times.

Cell culture apparatus for emulating gastrointestinal tract conditions and comprising at least two adjacent, microfluidic, cell cultivation channels separated by a permeable or semipermeable membrane, a first channel carrying gastrointestinal tract epithelial cells or tissues and a second channel carrying luminal and preferably mucosal microbiota, and wherein said second channel comprises one or more dwell chambers capable of providing a location for unattached luminal flora to reside away from any direct flow in said second channel, permits modelling of multiple sections of the gastrointestinal tract and control of retention times.

The invention provides a temperature sensitive cell culture surface and preparation method thereof. The preparation method comprises the following steps: (1) preparing a temperature sensitive primary liquid by adding a temperature sensitive compound and a free radical into a solvent, mixing and dissolving the same, and obtaining the temperature sensitive primary liquid; and (2) distributing the temperature sensitive primary liquid on a cell culture surface, and leaving the cell culture surface at 50-150° C. to react for 5-120 mins.

The invention provides a temperature sensitive cell culture surface and preparation method thereof. The preparation method comprises the following steps: (1) preparing a temperature sensitive primary liquid by adding a temperature sensitive compound and a free radical into a solvent, mixing and dissolving the same, and obtaining the temperature sensitive primary liquid; and (2) distributing the temperature sensitive primary liquid on a cell culture surface, and leaving the cell culture surface at 50-150° C. to react for 5-120 mins.

This invention relates to compositions of matter, methods, modules and instruments for automated mammalian cell growth and mammalian cell transduction followed by nucleic acid-guided nuclease editing in live mammalian cells. The present compositions and methods entail viral delivery of an editing cassette to live mammalian cells such that the editing cassettes edit the cells and the edited cells continue to grow, preferably using a fully-automated end-to-end instrument to process the cells without human intervention to enhance cell processing uniformity and to maintain the integrity of the cell culture.

This invention relates to compositions of matter, methods, modules and automated, end-to-end closed instruments for automated mammalian cell growth, reagent bundle creation and mammalian cell transfection followed by nucleic acid-guided nuclease editing in live mammalian cells.

A cell bag rotator is constructed to slowly agitate a plurality of cell bags by rotation along a rotation axis that is at least partially horizontal. The cell bag rotator comprises a plurality of plates to which a respective cell bag is attached. The cell bag rotator is suitable for mixing cells with particles for performing various types of subsequent cell therapy processes on the cells.

Provided herein are methods of culturing a mammalian cell and various methods that utilize these culturing methods. Also provided are multi-well cell culture plates, e.g., for use in perfusion culturing methods.