Recombinant production of growth factors for application in cell culture

Abstract

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 FGF-2, IGF-1, PDGF-BB and TGF-β1 in Escherichia coli. Using this expression system, we produced soluble GFs from species including bovine, chicken, and fish. Bioactivity analysis revealed orthologs with improved performance compared to commercially available alternatives. We stimated 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.

Employing active learning in the optimization of culture medium for mammalian cells

“…Generally, 10∼20% of FBS in the media was considered suitable for cell culture, as reported in enterocytes 39 , stem cells 40 , and hybridoma 41 . Such a reduced amount of FBS was supposed to be a preferred trend, considering the risk of contamination, the cost, and the batch-to-batch variability in cell production 42, 43 .…”

A Beefy-R culture medium: replacing albumin with rapeseed protein isolates

“…As such, it is likely that process scale-up can drive down costs substantially by leveraging economies of scale. Methods to optimize and enhance recombinant growth factors or overcome their requirement in cell culture could also dramatically reduce these costs 8,[30][31][32][33] . Additionally, during Beefy-9 development, it was shown that lowering the concentration of FGF-2 to 5 ng/mL did not drastically reduce growth, and so this simple strategy could reduce the cost of Beefy-R to $21.7/L.…”

The roles of growth factors and hormones in the regulation of muscle satellite cells for cultured meat production

“…Recombinant DNA technology can then be used to insert sequences of interest into bacterial systems. Using this strategy, active GFs, such as FGF-2, IGFs, PDGF-BB, and TGF-β1 were effectively developed in Escherichia coli, and soluble GFs from cattle, chickens, and fish were produced [130].…”