Cell Culture Media in Cultivated Meat - PDF

# Cell Culture Media in Cultivated Meat ## Overview Cell culture media is a major cost driver in cultivated meat production. - It accounts for **90-99% of production costs.** - Cost can be **over 300 euro per liter**, which translates to **1 euro per liter**. ## What is Culture Media? Cell cul...

# Cell Culture Media in Cultivated Meat ## Overview Cell culture media is a major cost driver in cultivated meat production. - It accounts for **90-99% of production costs.** - Cost can be **over 300 euro per liter**, which translates to **1 euro per liter**. ## What is Culture Media? Cell culture media is a blend of nutrients and components fed to cells to promote **growth and differentiation**. ## Components of Culture Media Cell media can be divided into two categories: - **Basal Media:** Contains basic building blocks and energy sources needed for cell survival and growth, including: - Glucose - Amino acids - Vitamins - Salts - Buffering agents which help maintain pH, osmolality, and cell homeostasis - **Added Factors:** Different types of molecules added to the media that are important for cell function, maintaining stemness, or steering differentiation. These are generally **cell and species-specific**. Examples include: - Fatty acids - Growth factors - Proteins - Vitamins - Buffering compounds - Inorganic salts - Carbon source (e.g. glucose) - Nitrogen source (e.g. amino acids) - Antioxidants - Small molecules - Surfactants - Anti-foam agents ## (Fetal) Animal Serum Animal serum is an *undefined mix of growth factors and other nutritive components* added as a supplement to basal media compositions. It was traditionally borrowed from medical research. - **Proliferation and Maintenance:** 20% FBS - **Differentiation:** 2% FBS However, animal serum has several drawbacks: - **High variability** - **Non-specific** - **High cost** - **Non-ethical** **Most companies in the cultivated meat industry have moved away from serum.** ## Serum-free Media for Muscle Cell Culture - **Research:** RNA sequencing during serum-starvation, differential gene expression before and after serum-starvation. - **Findings:** - **Surface receptors** _IGF1R, TFRC, LPAR1, OXTR, GCGR, CHRNA1_ are upregulated during serum-starvation in muscle cells. - **Solutions:** Supplementation with **ligands** such as Insulin, Transferrin, and **receptor-specific ligands** such as LPA, Oxytocin, and Glucagon shows promising results in promoting cell growth and differentiation. ## Serum-free Media for Fat Cell Culture - **Standard protocol:** 3-step adipogenic protocol using FBS-containing media. - **Drawbacks:** Insufficient differentiation across species. - **Simplified protocol: ** 1-step adipogenic protocol using FBS-free media. - **Advantages:** Robust differentiation across species. ## Media Cost Reduction ### Alternative Sourcing of Media Components - **Pharma grade to food-grade transition:** - Research findings: DMEM (Dulbecco's Modified Eagle's Medium) supplemented with food-grade (FG) basal media components supports growth of C2C12 (mouse myoblast) and bovine muscle-derived cells. However, serum is still needed. - **Hydrolysates and extracts:** - Research findings: Microalgae hydrolysates can be used as an alternative source of glucose and proteinogenic amino acids. However, they do not support differentiation. - **Agricultural and industrial sidestreams:** - Research is ongoing to find low-cost protein extracts and hydrolysates from plant-based agro-industrial waste. - **Protein isolates as replacement for growth factors:** - Research is ongoing to investigate the use of protein isolates as a cost-effective replacement for growth factors. ### Optimized Media Use - **Modelling:** - Research is ongoing to develop models to optimize media formulation. - **Improved feed strategies:** - Research is ongoing to investigate the use of improved feed strategies to reduce media costs. - **Cell line development:** - Research is ongoing to develop cell lines with faster proliferation and differentiation rates and minimal nutrient requirements. - **Media recycling and reuse:** - Research is ongoing to investigate the use of microalgae as a potential tool for recycling media components by removing ammonia and providing important nutrients for cell growth. ## Industry: Mosa Meat & Nutreco - **Mosa Meat:** - Signed an agreement with Nutreco to _reduce cost of cell feed and scale up production_. - **Nutreco:** - Opened a new facility dedicated to _food-grade cell feed for cultivated meat_. ## Industry: Mewery & SeaWith - **Mewery:** - Demonstrated _serum-free, microalgae-based growth medium_. - Received _pre-seed investment_ to advance their research. - **SeaWith:** - Cracked the code of _low-cost non-FBS growth media and scaffolding with microalgae_. ## Industry: Multus Media - **Machine learning:** - Using _AI to optimize cell culture media formulation_. ## Industry: Meatly & Believer Meats - **Meatly:** - Became the _first cultivated meat to be approved in Europe_. - _Protein-free media below 1 USD_. - **Believer Meats:** - _Serum-free media below 1 USD_. - _Production of cultivated chicken (50%) at 6/7 USD per kg_. ## Industry: ORF Genetics - **Production of recombinant-proteins in barley plants/seeds:** - _10-fold cost reduction when compared to other methods (e.g. E coli, yeast)_. The future of cultivated meat relies greatly on innovations and progress in media development. The industry is actively exploring cost-effective strategies to make this alternative protein source more accessible for a wider audience.