COMPOSITIONS AND METHODS FOR INCREASING THE EFFICIENCY OF CELL CULTURES USED FOR FOOD PRODUCTION
Provided herein are compositions and methods to make and use engineered cells, for the purpose of increasing the cell density of a culture comprising metazoan cells and for the production of a cultured edible product.
1 . A method for extending a renewal capacity of a cell culture comprising non-human cells having fibroblastic capacity, the method comprising:
a) introducing into the cells one or more polynucleotide sequences encoding a telomerase reverse transcriptase (TERT) or an engineered transcriptional activator capable of activating endogenous TERT expression; and
b) culturing the cells in a suspension cultivation infrastructure, adherent cultivation infrastructure, or a combination thereof.
2 . The method of claim 1 , further comprising modifying the cells to express one or more myogenic transcription factors, whereby expression of the one or more myogenic transcription factors differentiates at least a portion of the cells into cells having myogenic potential.
3 . The method of claim 2 , wherein the one or more myogenic transcription factors are selected from MYOD1, MYOG, MYF5, MYF6, PAX3, and PAX7.
4 . The method of claim 1 , wherein the cells are derived from poultry or a game species.
5 . The method of claim 1 , wherein the cells are derived from duck.
6 . The method of claim 1 , wherein the cells of step (a) comprise primary duck fibroblasts.
7 . The method of claim 1 , wherein the introducing step comprises ectopically expressing the polynucleotide sequence encoding TERT.
8 . The method of claim 1 , wherein the introducing step comprises transiently expressing the polynucleotide sequence encoding TERT.
9 . The method of claim 1 , wherein the introducing step comprises engineering the cells to stably express TERT.
10 . The method of claim 1 , wherein the TERT comprises activated TERT.
11 . A method for increasing insulin-like growth factor (IGF) concentration in a cell culture, comprising:
a) providing cells having myogenic or fibroblastic capacity; and
b) introducing into the cells one or more polynucleotide sequences encoding IGF,
wherein the concentration of IGF is increased relative to cells cultured without introduction of one or more polynucleotide sequences encoding IGF.
12 . The method of claim 11 , further comprising culturing the cells as a self-adherent aggregate.
13 . The method of claim 11 , further comprising culturing the cells in a suspension culture at a density of 50,000 to 1,000,000 cells per mL.
14 . The method of claim 11 , further comprising culturing the cells sequentially in both a suspension culture and an adherent culture.
15 . The method of claim 11 , wherein the introducing step comprises ectopically expressing the polynucleotide sequence encoding IGF.
16 . The method of claim 11 , wherein the introducing step comprises transiently expressing the polynucleotide sequence encoding IGF.
17 . The method of claim 11 , wherein the introducing step comprises engineering the cells to stably express IGF.
18 . The method of claim 11 , wherein the cells are derived from a poultry or a game species.
19 . The method of claim 11 , wherein the cells are derived from duck.
20 . The method of claim 11 , further comprising introducing into the cells one or more polynucleotide sequences encoding albumin, wherein the concentration of albumin is increased relative to cells cultured without introduction of one or more polynucleotide sequences encoding albumin.
21 . A method for decreasing ammonia concentration in a cell culture, comprising:
a) providing cells having myogenic or fibroblastic capacity; and
b) introducing into the cells one or more polynucleotide sequences encoding glutamine synthetase (GS),
wherein a concentration of ammonia is reduced relative to cells cultured without introduction of one or more polynucleotide sequences encoding GS.
22 . The method of claim 21 , further comprising culturing the cells as a self-adherent aggregate.
23 . The method of claim 21 , further comprising culturing the cells in a suspension culture at a density of 50,000 to 1,000,000 cells per mL.
24 . The method of claim 21 , further comprising culturing the cells sequentially in both a suspension culture and an adherent culture.
25 . The method of claim 21 , wherein the introducing step comprises ectopically expressing the polynucleotide sequence encoding GS.
26 . The method of claim 21 , wherein the introducing step comprises transiently expressing the polynucleotide sequence encoding GS.
27 . The method of claim 21 , wherein the introducing step comprises engineering the cells to stably express GS.
28 . The method of claim 21 , wherein the cells are derived from a poultry or a game species.
29 . The method of claim 21 , wherein the cells are derived from duck.
30 . The method of claim 21 , further comprising introducing into the cells one or more polynucleotide sequences encoding albumin; and wherein the concentration of albumin is increased relative to cells cultured without introduction of one or more polynucleotide sequences encoding albumin.