Produção sustentável em larga-escala de células de cavala para agricultura celular

One of the critical challenges of the growing cell-cultivated food industry is the ability to upscale production [1]. In fact, the scalable cultivation of fish cells in bioreactors is a complex endeavor that can slow down the introduction of these products in the market. This project proposes an integrated approach where scale-up studies are complemented by biological characterization of the cells and optimization of the culture medium. The project team’s combined expertise will allow them to tackle the expected challenges and bridge knowledge gaps to ensure a robust implementation of the work plan. The Project has 3 main objectives: 1) Scale-up of Mack1 cells using bioreactors; 2) Multi-Omic characterization of the Mack1 cell line during proliferation and myogenic differentiation supported by machine-learning algorithms; 3) Replacement of Fetal Bovine Serum by carotenoid-rich plant cell culture extracts. These objectives will be pursued on 3 different tasks, which will be interconnected. The Mack1 cell line, which was isolated by Saad and colleagues at the Kaplan Lab, is an immortalized Mackerel satellite cell line and is an outstanding starting material for cell-cultivated fish production. The authors already performed a preliminary characterization of these cells in terms of gene expression, but a more in-depth analysis is necessary to enable large-scale production and quality control. In particular, characteristic gene expression signatures of the satellite cell (or muscle stem cell) and myotube states, essential to assess cell identity and differentiation efficiency, are not clearly defined yet. This characterization will be made using bioinformatic techniques and a machine learning workflow developed by iBB and IT [2,3] which allows to analyze the whole transcriptome under different experimental conditions and time points. The second objective of the Project will address another major bottleneck of the cell-cultivated food field, the replacement of FBS by more sustainable and non-animal origin culture medium components. FBS is a high protein-containing mixture, widely used in animal cell culture media to promote cell survival and proliferation [4]. Albumin, the most abundant protein in FBS, has an important role as a carrier of a variety of lipids, amino acids, hormones, peptides and other molecules [5]. Our approach will be to obtain plant cell culture extracts, from the cell line Medicago A17 [6], and evaluate the possibility of replacing albumin with plant protein analogues. Besides albumin, there are other proteins present in FBS which have an important role in cell cultivation. In this sense, the team will collaborate with Defined Biosciences, a US-based company specialized in the production of culture medium components. Importantly, MACK4ALL will innovate using extracts from plant cell lines engineered to produce the carotenoids canthaxanthin and astaxanthin [7], which are expected to enhance the final product nutritionally and, due to their strong antioxidant properties, may potentially improve cell growth and viability, as well as minimize lipid oxidation which is an important cause of quality deterioration in seafood.