Recombinant production of growth factors for application in cell culture
Meenakshi Venkatesan,
Cameron Semper,
Stig Skrivergaard,
Rosa Di Leo,
Nathalie Mesa,
Martin Krøyer Rasmussen,
Jette Feveile Young,
Margrethe Therkildsen,
Peter J. Stogios,
Alexei Savchenko
Affiliations
Meenakshi Venkatesan
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E8, Canada
Cameron Semper
Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, AB T2N 4N1, Canada
Stig Skrivergaard
Department of Food Science, Aarhus University, 8200 Aarhus, Denmark
Rosa Di Leo
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E8, Canada
Nathalie Mesa
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E8, Canada
Martin Krøyer Rasmussen
Department of Food Science, Aarhus University, 8200 Aarhus, Denmark
Jette Feveile Young
Department of Food Science, Aarhus University, 8200 Aarhus, Denmark
Margrethe Therkildsen
Department of Food Science, Aarhus University, 8200 Aarhus, Denmark
Peter J. Stogios
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E8, Canada; Corresponding author
Alexei Savchenko
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E8, Canada; Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, AB T2N 4N1, Canada; Corresponding author
Summary: Culturing eukaryotic cells has widespread applications in research and industry, including the emerging field of cell-cultured meat production colloquially referred to as “cellular agriculture”. These applications are often restricted by the high cost of growth medium necessary for cell growth. Mitogenic protein growth factors (GFs) are essential components of growth medium and account for upwards of 90% of the total costs. Here, we present a set of expression constructs and a simplified protocol for recombinant production of functionally active GFs, including FGF2, IGF1, PDGF-BB, and TGF-β1 in Escherichia coli. Using this E. coli expression system, we produced soluble GF orthologs from species including bovine, chicken, and salmon. Bioactivity analysis revealed orthologs with improved performance compared to commercially available alternatives. We estimated that the production cost of GFs using our methodology will significantly reduce the cost of cell culture medium, facilitating low-cost protocols tailored for cultured meat production and tissue engineering.
