Upside Foods, Inc.

Provided herein are methods of engineering a cell line for reduced dependence on exogenous growth factor(s). In some embodiments, the method includes introducing into a cell one or more of: a polynucleotide comprising a coding sequence of a growth factor ligand; a polynucleotide comprising a coding sequence of a growth factor receptor; or a polynucleotide comprising a coding sequence of an activated downstream growth factor target, and culturing the cells in a cultivation infrastructure.

The present disclosure relates to systems, apparatuses, and methods for continuous cultivation and recovery of comestible non-human animal cells in a suspension bioreactor. For example, in one or more implementations, the disclosed methods include cultivating non-human animal cells in suspension culture within a bioreactor vessel, removing a measure of suspension culture from the bioreactor vessel while continuing to provide cell culture media thereto, and recovering a first portion of cultivated non-human animal cells from the removed measure of suspension culture while returning a second portion of cultivated non-human animal cells to the bioreactor vessel. Also, in some implementations, the disclosed systems include at least one sterile output module configured to prevent backflow contamination when recovering cultivated cells and removing spent media during continuous cultivation of non-human animal cells according to the disclosed methods.

Provided herein are methods of making a comestible cell-based meat composition where the method includes cooling the cell mass from a growth temperature to a temperature below a growth temperature; harvesting at least a portion of the cell mass; de-wetting the harvested cell mass with mechanically-generated pressure; forming the cell-based meat composition into a fillet; and optionally processing the fillet under high pressure, until the comestible cell-based meat composition satisfies a texture profile criteria.

The present disclosure relates to systems, apparatuses, and methods for preparing cell-based meat products (i.e., comestible meat products). In particular, a pipe-based bioreactor is disclosed having one or more substrates disposed therein. In one or more embodiments, the one or more substrates comprise a plurality of nominally spaced substrates conforming to an interior profile of an elongated enclosure of the pipe-based bioreactor. In some embodiments, multiple pipe-based bioreactors are interconnected a fluid source for preparing cell-based meat products. In addition, various methods and procedures for utilizing embodiments of pipe-based bioreactors are disclosed.

The present disclosure relates to systems, apparatuses, and methods for continuous cultivation and recovery of comestible non-human animal cells in a suspension bioreactor. For example, in one or more implementations, the disclosed methods include cultivating non-human animal cells in suspension culture within a bioreactor vessel, removing a measure of suspension culture from the bioreactor vessel while continuing to provide cell culture media thereto, and recovering a first portion of cultivated non-human animal cells from the removed measure of suspension culture while returning a second portion of cultivated non-human animal cells to the bioreactor vessel. Also, in some implementations, the disclosed systems include at least one sterile output module configured to prevent backflow contamination when recovering cultivated cells and removing spent media during continuous cultivation of non-human animal cells according to the disclosed methods.

Provided herein are methods of engineering a cell line for reduced dependence on exogenous growth factor(s). In some embodiments, the method includes introducing into a cell one or more of: a polynucleotide comprising a coding sequence of a growth factor ligand; a polynucleotide comprising a coding sequence of a growth factor receptor; or a polynucleotide comprising a coding sequence of an activated downstream growth factor target, and culturing the cells in a cultivation infrastructure.

Provided herein are transposon/transposase systems for transposing and selecting of transposed cells where the system includes a transposase or polynucleotide encoding a transposase; and an exogenous nucleic acid sequence comprising: a transposon comprising: a 5′ inverted terminal repeat (ITR), a first polynucleotide encoding a gene of interest; and a 3′ inverted terminal repeat (ITR); and (ii) a polynucleotide encoding an at least first selection marker located outside of the 5′ ITR and 3′ ITR, where upon introducing (a) and (b) into a cell the transposon is transposed into the cell's genome and the polynucleotide encoding the selection marker is episomal.

A plant fat-based scaffold for growing cell-based meat products for consumption. The scaffold comprises primarily plant fats or waxes in addition to cell binding proteins and optional additional components that assist in the growth of cultivated animal cells. The scaffold can exist in both a liquified state during sterilization and a solid state during the formation of the scaffold, the seeding of the cultivated cells, and the cellular growth phase. The scaffold is capable of remaining in the final product for consumption or is partially or completely melted out of the final product and recycled into raw material for forming new scaffolds.

Provided are methods and systems for producing a cell based product for dietary consumption, methods and systems for selecting cell lines with heat shock inducible genes associated with a fat or muscle phenotype, draw and fill methods and systems, methods and systems for a chemostat with cell retention, chicken or bovine cell lines, and edible bovine or avian cell lines obtained from the methods described herein.

A plant fat-based scaffold for growing cell-based meat products for consumption. The scaffold comprises primarily plant fats or waxes in addition to cell binding proteins and optional additional components that assist in the growth of cultivated animal cells. The scaffold can exist in both a liquified state during sterilization and a solid state during the formation of the scaffold, the seeding of the cultivated cells, and the cellular growth phase. The scaffold is capable of remaining in the final product for consumption or is partially or completely melted out of the final product and recycled into raw material for forming new scaffolds.