The cultivation medium is arguably the most essential component of modern cell cultivation technology. It provides the necessary support for cell survival, proliferation, and function outside of the animal body. In the context of cultivated meat, the medium is not just a nutritional solution, it is the biochemical environment that determines the success of cell expansion, differentiation, and ultimately, the quality of the final product. The ability to grow and study animal cells in vitro has revolutionized science and medicine over the past 150 years. One of the first documented experiments in this field dates back to the late 19th century, when Wilhelm Roux maintained neural plate cells from a chick embryo in saline solution for several days. A few years later, Montrose Burrows and Alexis Carrel developed the first long-term tissue cultivation system, cultivating animal cells using a medium enriched with animal plasma. Their cultures remained viable and proliferative for several months, marking a critical step in the development of cell cultivation as a scientific discipline. By the 1970s, large-scale applications of cell cultivation had emerged, most notably in the production of monoclonal antibodies. This milestone demonstrated the industrial potential of in vitro systems and laid the groundwork for biopharmaceutical manufacturing. Today, the same fundamental techniques are being adapted to an entirely new application: cultivated meat.
Despite the rapid progress in cell-based food technologies, cultivation media remain one of the major technical obstacles to large-scale commercialization. Animal cell cultivation media have been developed and optimized over the past century, but primarily for biomedical and pharmaceutical purposes. These industries typically produce high-value products in small quantities, where the cost of inputs is less critical. As a result, many traditional media formulations include expensive and animal-derived components, such as fetal bovine serum (FBS) or recombinant growth factors, which are neither economically feasible nor ethically ideal for food production. In contrast, cultivated meat demands a completely different set of priorities: food-grade safety, scalability, affordability, and public acceptability. This shift has prompted renewed efforts to reformulate media using non-animal, chemically defined, and cost-efficient alternatives that still meet the complex nutritional and signaling requirements of muscle, fat, and connective tissue cells.
Cultivation media can be broadly classified according to their composition and supplementation strategy. Serum-containing media rely on animal-derived supplements like FBS and are still commonly used in research. Serum-free media exclude serum but may contain purified proteins or hydrolysates. Protein-free media eliminate all proteinaceous supplements, relying on small molecules and synthetic components. In chemically defined media, all ingredients are known and quantified, offering superior reproducibility and safety. In the following sections, we will examine the composition of cultivation meda in more detail, explore the challenges associated with cost, sustainability, and performance, and discuss emerging strategies for creating media tailored specifically to the needs of cultivated meat production.
The formulation of cultivation media is central to the success of cell-based systems, including those used for cultivated meat. At its core, the medium must supply all the essential nutrients required for cellular survival and function. But beyond basic sustenance, it also serves as a signaling environment that can guide specific cellular behaviors such as proliferation, adhesion, migration, and differentiation into muscle, adipose, or connective tissue lineages. A comprehensive review of commercially available animal cell cultivation media is outlined in O’Neill et al. (2021).
To fulfill these roles effectively, the physical and chemical properties of the medium must closely mimic the physiological conditions within the body. Cultivation media are therefore built upon a saline solution and buffered to maintain physiological pH, typically around 7.4, with osmolality matched to the cytoplasmic environment of the target cell type. Deviations in pH or ion concentrations can significantly impair cell viability, making buffer systems a critical component of media design. In addition to their buffering roles, some salt components, such as calcium, magnesium, or potassium ions, also serve as secondary nutritional or signaling agents, influencing processes like cell adhesion, signal transduction, or membrane potential maintenance.
Beyond these physicochemical parameters, the success of cultivated cell systems also hinges on meeting the fundamental metabolic demands of the cells themselves. While each cell type, and indeed each species, presents its own set of metabolic preferences and requirements, most animal cells share a set of fundamental nutritional needs that can be addressed through general basic medium formulations. These requirements reflect the core biochemical demands of cellular life: energy generation, biosynthesis, redox balance, and maintenance of structural integrity. At the highest level, the nutritional inputs of cultivated cells can be categorized into two major groups: carbon sources and nitrogen sources. The uptake and metabolism of these macronutrients are tightly regulated by extracellular conditions, intracellular signaling pathways, and the proliferate or differentiative state of the cell.
Carbon-based compounds serve as the primary energy substrates and biosynthetic precursors for all cells. In the context of animal cell cultivation, carbohydrates (particularly glucose), selected amino acids, and in some cases fatty acids are dominant carbon donors. Despite the potential presence of other sugars (such as fructose, pyruvate, maltose, or sucrose) in the cultivation environment, animal cells are inherently optimized to metabolize glucose. Once taken up, glucose undergoes phosphorylation and enters a variety of metabolic pathways: glycolysis and subsequent fermentation or oxidative phosphorylation for ATP generation; the pentose phosphate pathway for nucleotide synthesis and redox homeostasis; and biosynthesis of fatty acids and amino acids. Alongside glucose, glutamine is an indispensable carbon source via glutaminolysis, contributing carbon atoms through conversion into α-ketoglutarate, which fuels the tricarboxylic acid cycle and supports anabolic metabolism. Although most animal cells are capable of synthesizing lipids de novo, the supplementation of select fatty acids in the medium can ease metabolic strain and enhance growth performance.
Crucially, oxygen, though not part of the medium per se, is a fundamental input in these pathways, particularly as the terminal electron acceptor in oxidative phosphorylation. Media systems must therefore ensure continuous and balanced oxygenation to support aerobic metabolism and energy efficiency. However, certain cell types, particularly satellite cells derived from muscle tissue, have demonstrated surprising adaptability to low-oxygen environments. For instance, mouse satellite cells have been shown to proliferate twice as rapidly under hypoxic conditions (2% O₂), and the myogenic potential of pig satellite cells was similarly enhanced at reduced oxygen levels.
Alongside carbon metabolism, nitrogen availability represents another fundamental requirement. Cellular structures such as proteins and nucleic acids are rich in nitrogen making glutamine (also a carbon donor) and other amino acids the primary nitrogen donors in media formulations. In rapidly dividing cells, nitrogen demand is particularly high, driven by anabolism associated with biomass expansion. Cells require both non-essential amino acids, which the body can synthesize, and essential amino acids, which must be obtained from external sources. Among essential amino acids for most animal cells are: histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. Glutamine is especially critical, not only as a nitrogen donor, but also for NADPH regeneration, a cofactor essential for numerous biosynthetic reactions. However, glutaminolysis can lead to ammonia accumulation in the medium, a cytotoxic byproduct that must be monitored or mitigated. Potentially through genetic modification, cells can synthesize glutamine endogenously.
In addition to these macro-elements, cells require a carefully balanced composition of vitamins and minerals, though their specific roles and concentrations vary across cell types and functional states. Water-soluble vitamins, notably the B-complex and vitamin C, are widely considered essential. Vitamin C supports collagen synthesis by fibroblasts, contributing to the structural properties of cultivated tissue. Fat-soluble vitamins (such as A, D, E, and K) may be beneficial under specific conditions. For example, vitamin E is antioxidant and vitamin A promotes myogenesis though their routine inclusion is debated.
Minerals play diverse roles in media performance. Some act as osmolytes (e.g., sodium, potassium, bicarbonate). Others are enzyme cofactors (e.g., magnesium, iron, zinc) or are integrated into cellular structures and signaling pathways (e.g., calcium, phosphate, sulfate). Trace elements such as copper, iodine, selenium, molybdenum, and manganese are also often added in micro-molar concentrations to support enzymatic function and proper cellular function.
To actively guide cells through desired developmental pathways, such as proliferation, differentiation, or migration, cultivation media must contain growth factors, a group of signaling proteins that mimic the body’s natural regulatory cues. By binding to specific cell surface receptors, they initiate intracellular signaling cascades that regulate cell survival, proliferation, migration, and differentiation. In the context of cultivated meat, these molecules are added to the cultivation medium to mimic the natural cues that would ordinarily come from an animal’s body, thereby guiding cells through the stages necessary to form muscle, fat, or connective tissue. Key examples include insulin-like growth factors (IGF-1 and IGF-2), transforming growth factor beta (TGF-β), fibroblast growth factors (such as FGF-2 and FGF-21), platelet-derived growth factor (PDGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF). Each plays a specific role: FGFs, for instance, are critical in regulating proliferation and tissue regeneration, while PDGFs are potent mitogens that drive cell division. Despite their functional importance, growth factors remain one of the most expensive components. As a result, considerable effort has been directed toward reducing their cost and improving their scalability. One of the most promising strategies is the use of recombinant DNA technology to produce these proteins more affordable. For example, growth factors such as FGF-2, IGFs, PDGF-BB, and TGF-β1 have been successfully produced in microbial systems like Escherichia coli. However, optimizing the composition of growth factors remains a challenge. Different cell types require specific signaling environments, and the ideal balance of growth factors for promoting proliferation versus differentiation can vary not only between species, but even between cell lines of the same tissue type. Scaling up these complex combinations from lab-scale to industrial-scale bioreactors without compromising performance is a key hurdle for the field.
Hormones, similarly, are powerful endocrine signaling molecules that play a pivotal role in regulating cellular processes such as gene expression, metabolism, growth, and differentiation. Their activity is mediated through specific receptors located on the cell membrane or within the cytoplasm, where hormone binding triggers complex intracellular signaling cascades. In cultivated meat production, hormones may be added to the cultivation medium to guide cells towards desired behaviors, such as proliferation, differentiation, and hypertrophy, depending on the target tissue type. Many hormones naturally found in FBS contribute to cell survival and function. These include insulin, cortisol, growth hormone, parathyroid hormone, thyroid hormones (T3 and T4), follicle-stimulating hormone, prolactin, testosterone, and progesterone. Each plays a distinct role. Insulin supports cell survival and self-renew. Testosterone and other androgens promote muscle growth. Cortisol helps regulate energy homeostasis and metabolism. Thyroid hormones stimulate cell proliferation and differentiation and are important for muscle recovery and myogenesis. Because some hormones act only on specific cell types, their use must be carefully tailored to the needs of the cultivated cells. For example, myoblasts (muscle precursor cells) respond to androgens and thyroid hormones during the differentiation process, while fibroblasts may be more influenced by corticosteroids or insulin. However, since cultivated meat is intended for human or animal consumption, the inclusion of hormones in the production process raises critical questions, particularly in light of widespread public concern over hormonal residues in food. Hormones in the diet are often perceived as undesirable, and regulatory frameworks in many regions place strict limits on their use. As a result, the cultivated meat industry is increasingly shifting away from hormone supplementation, striving instead to develop hormone-free strategies that still support efficient cell growth and differentiation while ensuring food safety and consumer acceptance.
In conventional biomedical research and laboratory scale cell cultivation, antibiotics (penicillin, streptomycin, amphotericin B, and gentamicin) are frequently included in media to prevent bacterial contamination. While not essential for cell growth, they offer a level of convenience by protecting cultures. However, their presence comes at cost. Antibiotics can interfere with metabolism of sensitive cell types, potentially affecting proliferation, differentiation, and viability. Moreover, their use may mask underlying contamination with mycoplasma or antibiotic-resistant bacteria, compromising safety and reproducibility of the cultivation system. In research settings, these risks are often tolerated and carefully monitored. In food production, however, they present a more serious concern. In the context of cultivated meat, the use of antibiotics in the media raises health and ethical questions. Residual antibiotics in the final product could pose a risk to consumers with hypersensitivities or allergies to specific antibiotics. More broadly, the inclusion of antibiotics in large-scale food production may contribute to the growing global problem of antimicrobial resistance. From a consumer acceptance standpoint, the presence of antibiotics, even in trace amounts, may hinder public trust and undermine the perceived safety of cultivated meat products. Therefore, the cultivated meat industry is actively working toward antibiotic-free cultivation systems. This involves strict aseptic processing, closed bioreactors, rigorous monitoring protocols, and improved media formulations that minimize the need for antimicrobial intervention. Achieving reliable contamination control without antibiotics is not only a technical challenge, but a prerequisite for regulatory approval and consumer confidence.
A significant milestone in the development of cell cultivation technology came in the 1950s with the introduction of fetal bovine serum as a standard supplement in cultivation media. Originally intended to stimulate cell growth, FBS was soon recognized as an essential and multifunctional component, providing a complex mixture of nutrients and bioactive molecules necessary for cell attachment, proliferation, and long-term maintenance. Its composition includes a wide array of substances such as albumin, fetuin, hormones, vitamins, trace elements, growth factors, cytokines, and adhesion molecules. In total FBS contains approximately 1800 proteins and over 4000 metabolites, making it a potent, albeit largely undefined, biological supplement.
FBS is derived from the blood of bovine fetuses collected from pregnant cows during routine slaughterhouse procedures, which has raised ethical concerns. More than two million bovine fetuses are used globally each year to produce an estimated 800,000 liters of FBS. This practice not only raises questions about animal welfare, but also introduces significant technical and practical limitations. FBS is inherently variable between batches, difficult to standardize, and carries risk of contamination with infectious agents such as viruses or prions. From the perspective of cultivated meat, such inconsistencies can translate directly into changes in muscle cell phenotype, ultimately impacting the sensory qualities and reproducibility of the final product. In the context of cultivated meat, reliance on FBS or other animal-derived ingredients is not viable. The media must be food-grade, cost-effective, and safe for large-scale manufacturing. It must also be consistent, scalable, and free of animal origin components to meet both ethical and regulatory expectations. For these reasons, the elimination of serum is one of the central challenges in the formulation of cultivation media for meat production.
It is important to note that nearly all established cell lines in research and industry were originally developed using serum-containing media and later adapted to serum-free formulations. This transition requires gradual and carefully controlled adaptation steps, as abrupt shifts in nutrient or signaling environments can lead to reduced proliferation, altered cell behavior, or complete cultivation failure. Ultimately, the development of fully defined, serum-free, and animal-free media tailored specifically for cultivated meat remains a critical focus in the field. Success in this area will not only improve scalability and cost-efficiency but also that cultivated meat meets the safety, regulatory and ethical expectations of both producers and consumers.
A number of serum-free media have already been developed for biopharmaceutical and biomedical research, often relying on recombinant growth factors, such as insulin, fibroblast growth factor 2 (FGF2), and transforming growth factor beta (TGF-β) . One such formulation is Essential 8™, a defined, xeno-free medium supplemented with the aforementioned growth factors and additional vitamins, minerals, and buffering agents. Similarly, Fibroblast Growth Medium™ has been optimized specifically for the cultivation of human fibroblasts. However, these formulations are prohibitively expensive and not designed with food production in mind. To address the cost barrier, alternative strategies are under active development. One promising avenue is the use of genetically modified cell lines that are capable of endogenously producing essential growth factors, thus eliminating the need for repeated supplementation. Another approach involves utilizing human platelet lysates, derived as a by-product from conventional blood donations. These lysates are rich in growth factors and cytokines and have shown potential for supporting cell proliferation and attachment. However, their use in cultivated meat faces obstacles related to cross-species compatibility, regulatory classification, and supply logistics.
In the search for acceptable alternatives to serum, hydrolysates have gained increasing attention. These complex mixtures, derived from enzymatic, acidic, alkaline, or fermentative breakdown of proteins, offer a rich and diverse source of nutrients for animal cells. Hydrolyzates can be produced from a wide range of raw materials, both animal (such as chicken, pork, and fish) and plant-based (including soy, pea, rice, or rapeseed). Their composition typically includes peptides, free amino acids, minerals, carbohydrates, lipids, and residual proteins that reflect the biochemical makeup of the originate substrate. Hydrolysates were first tested as serum replacement in the late 1970s, initially using products derived from chicken and fish. However, these animal-based versions presented similar safety, ethical, and variability concerns as serum itself. In response, attention has gradually shifted towards plant-based hydrolysates, which are more suitable for applications in food production and align with principles of animal-free cultivation.
The advantages of hydrolysates are manifold. They are relatively inexpensive, widely available, and have demonstrated beneficial effects beyond basic nutrition, including the promotion of cell proliferation, viability, and stress resistance. This makes them notably attractive for large-scale application such as cultivated meat production, where cost and consistency are critical. However, hydrolysates also come with significant challenges. Unlike chemically defined media, their extract composition is often unknown or insufficiently characterized. Their nutritional profile and bioactivity are influenced by several factors, including the type and quality of the raw material, the hydrolysis method employed, and the degree of breakdown achieved. As a result, variability between batches can be substantial, leading to inconsistencies in cell performance. Additionally, since hydrolysates are processed products, contaminants can be introduced either from the source material or during the hydrolysis process itself. These contaminants may negatively affect cell growth or compromise the sterility of the medium, especially serious concern in sensitive and high-purity systems such as those used in cultivated meat production.
Although more research is needed to fully understand the effects of hydrolysate components on specific cell types, early studies suggest promising potential. For example, Tuomisto & Teixeira de Mattos (2011) reported the successful use of cyanobacterial hydrolysates as a media supplement for muscle cells. Such findings underscore the importance of continuing to explore and optimize hydrolysate formulations, particularly those based on food-grade, plant-derived sources. Ultimately, while hydrolysates are not fully refined replacements for serum, they represent an important step toward affordable and functional media solutions. Future progress in their standardization, purification, and mechanistic understanding could unlock their broader application.
Despite significant advancement, the design of cultivation media that meet the unique demands of cultivated meat production, namely scalability, affordability, and product quality, remains a complex and unresolved challenge. Beyond meeting the basic metabolic needs of cells, the medium must also account for stability, sterility, and consumption rates of its components. Many biologically active ingredients, such as growth factors and vitamins, are heat-sensitive and cannot withstand conventional sterilization processes. This necessitates the use of alternative sterilization methods, including filtration and irradiation, or development of heat-stable formulations tailored to the manufacturing environment. Interestingly, the composition of the medium may also influence the sensory attributes of the final product. Although this area remains underexplored, the accumulation of certain compounds (such as glutamic acid or asparagine) within cultivated cells could impact taste contributing to umami or other flavor profiles. As the cultivated meat industry evolves from laboratory to consumer-facing products, understanding and managing such sensory effects will become increasingly important. Moreover, no single universal medium is likely to satisfy all applications. Different cell types have distinct nutritional and signaling requirements, and these requirements can shift dramatically between the proliferation and differentiation phases. Media must therefore be tailored not only to the cell type, but also to the developmental stage of the cells. Ultimately, the development of next-generation media for cultivated meat represents one of the most critical areas for innovation in the field. It is where biology, chemistry, engineering, and food science converge, with the potential to redefine how we nourish the world.
Ahmad, S. S.; Chun, H. J.; Ahmad, K.; Shaikh, S.; Lim, J. H.; Ali, S.; Han, S. S.; Hur, S. J.; Sohn, J. H.; Lee, E. J.; Choi, I. The roles of growth factors and hormones in the regulation of muscle satellite cells for cultured meat production. Journal of Animal Science and Technology, 2023, 65, 1, 16-31. https://doi.org/10.5187/jast.2022.e114
Arora, M. Cell Culture Media: A Review. Labome, 2013, 3, 75, 24. https://dx.doi.org/10.13070/mm.en.3.175
Giglio, F.; Scieuzo, C.; Ouazri, S.; Pucciarelli, V.; Ianniciello, D.; Letcher, S.; Salvia, R.; Laginestra, A.; Kaplan, D. L.; Falabella, P. A Glance into the Near Future: Cultivated Meat from Mammalian and Insect Cells. Small Science, 2024, 4, 2400122. https://doi.org/10.1002/smsc.202400122
Ho, Y. Y.; Lu, H. K.; Lim, Z. F. S.; Lim, H. W.; Ho, Y. S.; Ng, S. K. Applications and analysis of hydrolysates in animal cell culture. Bioresources and Bioprocessing, 2021, 8, 93. https://doi.org/10.1186/s40643-021-00443-w
Chelladurai, K. S.; Christyraj, J. D. S.; Rajagopalan, K.; Yesudhason, B. V.; Venkatachalam, S.; Mohan, M.; Vasantha, N. Ch.; Christyraj, J. R. S. S. Alternative to FBS in animal cell culture – An overview and future perspective. Heliyon, 2021, e07686. https://doi.org/10.1016/j.heliyon.2021.e07686
Lee, S. Y.; Lee, D. Y.; Yun, S. H.; Lee, J.; Mariano, E. Jr.; Park, J.; Choi, Y.; Han, D.; Kim, J. S.; Hur, S. J. Current technology and industrialization status of cell-cultivated meat. Journal of Animal Science and Technology, 2024, 66, 1, 1-30. https://doi.org/10.5187/jast.2023.e107
O’Neill, E. N.; Cosenza, Z. A.; Baar, K.; Block, D. E. Considerations for the development of cost-effective cell culture media for cultivated meat production. Comprehensive Reviews in Food Science and Food Safety, 2021, 1-24. https://doi.org/10.1111/1541-4337.12678
Tuomisto, H. L.; Teixeira de Mattos, M. J. Environmental impacts of cultured meat production. Environmental Science and Technology, 2011, 45, 6117–6123. https://doi.org/10.1021/es200130u
Yao, T.; Asayama, Y. Animal-cell culture media: History, characteristics, and current issues. Reproductive Medicine and Biology, 2017, 21, 16, 2, 99-117. https://doi.org/10.1002/rmb2.12024
The cultivation medium is one of the most important building blocks of modern cell cultivation. In simple terms, it is the “liquid environment” that keeps cells alive outside an animal’s body. It provides cells with energy, raw materials, and the biochemical signals they need to grow and function. In cultivated meat, the medium is far more than a basic nutrient mix: it helps determine whether cells expand efficiently, whether they mature into muscle or fat, and how good the final product can become. The idea of keeping animal cells alive outside the body has a long history. In the late 19th century, scientists already managed to maintain chick embryo cells in simple saline solutions. Soon after, early tissue culture pioneers developed more complex systems using animal plasma, allowing cells to survive and proliferate much longer. Over the decades, cell cultivation became a cornerstone of medicine and biotechnology, enabling major breakthroughs such as large-scale production of monoclonal antibodies in the 1970s. Today, many of the same core principles are being adapted for a new purpose: producing cultivated meat.
Despite rapid progress in cell-based food technologies, cultivation media remain one of the biggest barriers to bringing cultivated meat to market at scale. Most existing media were designed for biomedical and pharmaceutical applications, where products are high-value and produced in relatively small quantities. In those settings, expensive ingredients are often acceptable. Many traditional media also rely on animal-derived components, especially fetal bovine serum (FBS), or on costly purified growth factors. For cultivated meat, the priorities are different: the medium must be food-safe, affordable, scalable, and widely acceptable to the public. That is why the field is investing heavily in developing animal-free, more consistent, and cost-efficient media that still meet the complex needs of muscle, fat, and connective tissue cells.
Cultivation media are commonly grouped based on what they contain and how they are supplemented. Serum-containing media include animal-derived supplements like FBS and are still widely used in research because they support many cell types reliably. Serum-free media remove serum, but may still contain purified proteins or complex nutrient blends such as hydrolysates. Protein-free media go further and avoid protein supplements altogether, relying mainly on small molecules and other defined ingredients. Chemically defined media represent the most controlled category: every component is known and measured, which improves reproducibility and safety. For cultivated meat, moving toward animal-free and more defined media is a central goal, but it remains technically and economically challenging.
A successful medium has to do two jobs at once. First, it must provide the basic nutrients cells need to survive and grow. Second, it must act as a “communication platform” that nudges cells toward specific behaviors, such as rapid growth, attachment, migration, or differentiation into muscle or fat. In other words, the medium is both food and instruction manual for the cells.
To work properly, media also need to mimic key aspects of the internal environment of the body. They are built on a salt solution and buffered to maintain a stable pH, usually around 7.4. Osmolality is also adjusted so cells do not swell or shrink from imbalanced fluid pressure. Even small deviations in pH or ion levels can damage cells, which is why buffering systems are essential. Some salts and ions, such as calcium, magnesium, and potassium, do more than stabilize chemistry: they can also influence cell adhesion, signaling, and electrical balance across membranes.
At a broader level, most cells share common nutritional needs that can be approached through general formulations. These needs reflect the basics of cellular life: energy production, building cellular components, and maintaining redox balance. For simplicity, the major nutritional inputs can be viewed as carbon sources and nitrogen sources, even though in practice many ingredients contribute to both.
Carbon sources provide energy and supply building blocks for growth. Animal cells are especially optimized to use glucose. Once cells take up glucose, they can break it down for energy and route it into pathways that support growth: producing ATP (the cell’s energy currency), generating molecules needed for DNA and RNA, and creating precursors for fats and amino acids. Alongside glucose, glutamine is another major contributor. It supports energy metabolism through pathways that feed into the cell’s central energy cycle and also helps supply materials for biosynthesis. Some media may also include selected fatty acids, not necessarily because cells cannot make fats themselves, but because supplying them can reduce metabolic strain and improve performance.
Oxygen matters here as well. Even though it is not a medium ingredient in the usual sense, it is essential for efficient energy generation in many cell types. In cultivated meat systems, oxygen supply must be managed carefully, especially at scale. Interestingly, some muscle-related cells can grow well under lower oxygen conditions, and in certain cases their proliferation or muscle-forming potential may even improve.
Nitrogen is essential because proteins and nucleic acids are nitrogen-rich. Amino acids are therefore key ingredients, serving as both building blocks and nitrogen donors. Cells require a mixture of non-essential amino acids (which they can synthesize) and essential amino acids (which must be provided). Essential amino acids include histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. Glutamine is especially important here too, because it contributes nitrogen as well as carbon and supports the production of molecules needed for biosynthesis. However, heavy glutamine use can lead to ammonia accumulation, which can harm cells if it builds up. This is one reason why medium design also has to consider byproducts and waste, not only nutrients.
Beyond macronutrients, cells depend on vitamins and minerals to support enzyme function, signaling, and structural stability. Water-soluble vitamins, especially the B vitamins and vitamin C, are widely regarded as essential in most systems. Vitamin C is particularly relevant because it supports collagen synthesis, which can influence structural properties in tissue formation. Fat-soluble vitamins such as A, D, E, and K may be useful in some contexts, but their routine inclusion depends on the cell type and the goals of cultivation.
Minerals have multiple roles: some help set osmotic balance, others act as cofactors for enzymes (like magnesium, iron, and zinc), and some contribute directly to signaling and structural functions (such as calcium and phosphate). Trace elements (such as copper, iodine, selenium, molybdenum, and manganese) are typically added in very small concentrations but can still be important for healthy cell function.
Nutrients alone are not enough. To guide cells toward specific behaviors (rapid growth, survival, migration, or differentiation( media often include growth factors. These are signaling proteins that act like “biological instructions,” binding to receptors on the cell surface and triggering internal programs. In cultivated meat development, growth factors help mimic the cues cells would receive in an animal body. Examples include insulin-like growth factors (IGF-1 and IGF-2), transforming growth factor beta (TGF-β), fibroblast growth factors (such as FGF-2 and FGF-21), platelet-derived growth factor (PDGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF). Each plays a different role, and the right combination depends on the cell type and whether the goal is expansion or maturation. A key challenge is cost. Growth factors are often among the most expensive medium components. One strategy for reducing cost is producing them using recombinant methods in microbial systems such as E. coli. Even with improved production, formulating the “right recipe” remains difficult because different cells, and different stages of the same cell, need different signaling environments.
Hormones can also shape cell behavior by influencing gene expression, metabolism, and differentiation. Many hormones present in serum contribute to cell survival and growth, including insulin, cortisol, thyroid hormones, growth hormone, testosterone, and others. In cell cultivation, hormones may be used to support proliferation, maturation, or muscle growth processes. However, cultivated meat is intended for consumption, and hormones are a sensitive topic for the public. Many people are concerned about hormones in food, and regulations in many regions strictly limit their use. Because of this, the cultivated meat field is increasingly shifting toward strategies that eliminate hormone supplementation, aiming to maintain efficiency while improving consumer trust and regulatory compatibility.
In laboratory research, antibiotics are often added to media as a protective measure against contamination. This can be useful at a small scale, but it is not ideal for food production. Antibiotics can influence cell behavior, mask low-level contamination, and create risks linked to antibiotic resistance. In the cultivated meat context, even the possibility of residues can reduce consumer confidence, and large-scale antibiotic use would raise ethical and public health concerns. For these reasons, cultivated meat developers are working toward antibiotic-free cultivation. That requires strict aseptic processing, closed production systems, strong monitoring protocols, and robust process design so contamination prevention is achieved through engineering and hygiene rather than antimicrobial additives.
Fetal bovine serum became a common supplement in the 1950s because it strongly supports cell growth. It contains a highly complex mixture of proteins, nutrients, hormones, trace elements, growth factors, and adhesion molecules. This complexity is exactly why it works so well, but it is also why it is problematic. Serum is biologically variable from batch to batch, making it difficult to standardize and potentially affecting cell behavior and product consistency. It also carries contamination risks.
The ethical concerns are equally significant. FBS is collected from bovine fetuses during slaughterhouse procedures, and global use involves millions of fetuses each year. For cultivated meat, which is often promoted partly for its ethical advantages, reliance on FBS is widely viewed as incompatible with the long-term vision. As a result, removing serum is one of the biggest priorities in medium development. It is also technically challenging, because many cell lines were originally developed in serum-containing conditions and must be adapted gradually to serum-free environments.
Serum-free media already exist for biomedical applications, often relying on purified recombinant growth factors and carefully optimized compositions. Examples include defined xeno-free formulations developed for human cells. These systems demonstrate what is possible, but they are typically far too expensive for food production. Several alternative strategies are being explored. One approach is developing cell lines that produce some required growth signals themselves, reducing the need for repeated supplementation. Another is using platelet lysates, by-products of blood donations, which contain growth factors and can support cell growth, though this route raises questions about classification, cross-species compatibility, and supply.
Hydrolysates are also receiving strong interest. These nutrient-rich mixtures are created by breaking down proteins into peptides and amino acids. They can be sourced from animal or plant materials, but plant-based hydrolysates are especially attractive for cultivated meat because they better align with safety and ethical goals. Hydrolysates can support growth and resilience and are relatively affordable, but they also come with drawbacks: their composition is complex and not always fully characterized, batch variability can be high, and there is potential for contaminants depending on sourcing and processing. Still, they represent an important bridge between expensive fully defined systems and the affordability required for large-scale food manufacturing.
Even with major advances, designing cultivation media that meet the specific demands of cultivated meat (low cost, scalability, safety, and consistent performance) remains one of the field’s hardest problems. Media must not only nourish cells but also remain stable, sterile, and compatible with industrial production. Many bioactive ingredients are heat-sensitive, meaning standard sterilization methods are not always possible and alternative approaches may be required. The medium may also influence product quality in unexpected ways, including potential effects on taste-related compounds. And there is unlikely to be one universal recipe: different cell types and different cultivation stages (growth vs. differentiation) require different nutrient and signaling profiles. For all these reasons, medium development is one of the most critical innovation frontiers in cultivated meat, where biology, chemistry, engineering, and food science come together to shape what will be possible at scale.
Ahmad, S. S.; Chun, H. J.; Ahmad, K.; Shaikh, S.; Lim, J. H.; Ali, S.; Han, S. S.; Hur, S. J.; Sohn, J. H.; Lee, E. J.; Choi, I. The roles of growth factors and hormones in the regulation of muscle satellite cells for cultured meat production. Journal of Animal Science and Technology, 2023, 65, 1, 16-31. https://doi.org/10.5187/jast.2022.e114
Arora, M. Cell Culture Media: A Review. Labome, 2013, 3, 75, 24. https://dx.doi.org/10.13070/mm.en.3.175
Giglio, F.; Scieuzo, C.; Ouazri, S.; Pucciarelli, V.; Ianniciello, D.; Letcher, S.; Salvia, R.; Laginestra, A.; Kaplan, D. L.; Falabella, P. A Glance into the Near Future: Cultivated Meat from Mammalian and Insect Cells. Small Science, 2024, 4, 2400122. https://doi.org/10.1002/smsc.202400122
Ho, Y. Y.; Lu, H. K.; Lim, Z. F. S.; Lim, H. W.; Ho, Y. S.; Ng, S. K. Applications and analysis of hydrolysates in animal cell culture. Bioresources and Bioprocessing, 2021, 8, 93. https://doi.org/10.1186/s40643-021-00443-w
Chelladurai, K. S.; Christyraj, J. D. S.; Rajagopalan, K.; Yesudhason, B. V.; Venkatachalam, S.; Mohan, M.; Vasantha, N. Ch.; Christyraj, J. R. S. S. Alternative to FBS in animal cell culture – An overview and future perspective. Heliyon, 2021, e07686. https://doi.org/10.1016/j.heliyon.2021.e07686
Lee, S. Y.; Lee, D. Y.; Yun, S. H.; Lee, J.; Mariano, E. Jr.; Park, J.; Choi, Y.; Han, D.; Kim, J. S.; Hur, S. J. Current technology and industrialization status of cell-cultivated meat. Journal of Animal Science and Technology, 2024, 66, 1, 1-30. https://doi.org/10.5187/jast.2023.e107
O’Neill, E. N.; Cosenza, Z. A.; Baar, K.; Block, D. E. Considerations for the development of cost-effective cell culture media for cultivated meat production. Comprehensive Reviews in Food Science and Food Safety, 2021, 1-24. https://doi.org/10.1111/1541-4337.12678
Tuomisto, H. L.; Teixeira de Mattos, M. J. Environmental impacts of cultured meat production. Environmental Science and Technology, 2011, 45, 6117–6123. https://doi.org/10.1021/es200130u
Yao, T.; Asayama, Y. Animal-cell culture media: History, characteristics, and current issues. Reproductive Medicine and Biology, 2017, 21, 16, 2, 99-117. https://doi.org/10.1002/rmb2.12024
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