Engineering cell lines capable of proliferation in growth factor free media formulations
AssigneeUpside Foods, Inc.
InventorsSukhdeep Singh Dhadwar, Rachel Michele Schumaker
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Abstract
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.
Claims22
A method for culturing an avian cell line in a cell culture media, said method comprising:
- (a) seeding an edible cell culture media with an edible, avian cell line arising from cells transduced with
- a polynucleotide comprising a single coding sequence encoding a single growth factor receptor or growth factor, which is an FGFR, a PDGFR, an activated FGFR, or an activated PDGFR, or a combination of solely a coding sequence encoding an FGFR or an activated FGFR and a coding sequence encoding an IGF;
- wherein the polynucleotide is engineered to express the coding sequence or the combination of coding sequences in the cell line;
- and
- (b) culturing the cell line in the cell culture media substantially free of exogenous growth factors,
- wherein the cell line proliferates in the absence of exogenous growth factors over 72 hours of culture to a cell density equivalent to a cell density over 72 hours of culture time of a comparable cell line not comprising the polynucleotide and cultured in the presence of the exogenous growth factors.
A method for culturing an edible, avian cell line in a cell culture media, said method comprising:
- (a) seeding an edible cell culture media with an edible, avian cell line arising from cells transduced with
- a polynucleotide comprising a combination of solely a coding sequence encoding IGF1 and a coding sequence encoding FGF;
- wherein the polynucleotide is engineered to express the coding sequences in the cell line;
- and
- (b) culturing the cell line in the cell culture media substantially free of exogenous growth factors,
- wherein the cell line proliferates in the absence of the exogenous growth factors over 72 hours of culture to a cell density equivalent to a cell density over 72 hours of culture time of a comparable cell line not comprising the polynucleotide and cultured in the presence of exogenous IGF1 and FGF.
A method for culturing an edible, avian cell line in a cell culture media, said method comprising:
- (a) seeding an edible cell culture media with an edible, avian cell line arising from cells transduced with
- a polynucleotide comprising a single coding sequence of a single growth factor, which encodes FGF2 or FGF3;
- wherein the polynucleotide is engineered to express the coding sequence in the cell line;
- and
- (b) culturing the cell line in the cell culture media substantially free of exogenous growth factors, wherein the cell line is capable of proliferating to a cell density higher than the seeded cell density over 72 hours of culture time of a comparable cell line not comprising the polynucleotide.
Dependencies are inferred from the claim text.