Analysis of current technology status for the industrialization of cultured meat.

Cultured meat is expected to become an important material for future food production; however, contrary to initial expectations, the full-scale industrialization of cultured meat is slow and the actual level and opened technology amount is very limited. This study reviews the publicly available technologies of cultured meat and suggests future developmental directions and research agenda. As a result of analyzing papers, patents, and press releases published over the past 10 years, it was found that cultured meat production technology is still at the prototype production level. This is because most papers published are about culture medium and scaffold development, culture conditions, and there is almost no research on finished cultured meat products. Worldwide, most of the filed patents are for producing cultured meat principles; most of them do not use food-grade materials and are not economically feasible for industrialization. Therefore, future research on the industrialization of cultured meat should focus on effective acquisition technologies for satellite cells; cell lineage and undifferentiated state maintenance technologies; the development of serum-free media and culture devices; the prevention of genetic modification, safety verification, and mass production. Furthermore, basic research on mechanisms and influencing factors related to cultured meat production is warranted.

Review of technology and materials for the development of cultured meat.

Cultured meat production technology suggested that can solve the problems of traditional meat production such as inadequate breeding environment, wastewater, methane gas generation, and animal ethics issues. Complementing cultured meat production methods, sales and safety concerns will make the use of cultured meat technology easier. This review contextualizes the commercialization status of cultured meat and the latest technologies and challenges associated with its production. Investigation was conducted on materials and basic cell culture technique for cultured meat culture is presented. The development of optimal cultured meat technology through these studies will be an innovative leap in food technology. The process of obtaining cells from animal muscle, culturing cells, and growing cells into meat are the basic processes of cultured meat production. The substances needed to production of cultured meat were antibiotics, digestive enzymes, basal media, serum or growth factors. Although muscle cells have been produced closer to meat due to the application of scaffolds materials and 3 D printing technology, still a limit to reducing production costs enough to be used as foods. In addition, developing edible materials is also a challenge because the materials used to produce cultured meat are still not suitable for food sources.

Subacute feeding toxicity of low-sodium sausages manufactured with sodium substitutes and biopolymer-encapsulated saltwort (Salicornia herbacea) in a mouse model.

BACKGROUND: Low-sodium sausages were manufactured using sodium substitution and biopolymer encapsulation. A diet comprising 10% treatment sausages (six treatment groups: C (100% NaCl), T1 (55% sodium substitute + 45% saltwort salt), T2 (55% sodium substitute + 45% saltwort salt with chitosan), T3 (55% sodium substitute + 45% saltwort salt with cellulose), T4 (55% sodium substitute + 45% saltwort salt with dextrin), and T5 (55% sodium substitute + 45% saltwort salt with pectin)) was added to a 90% commercial mouse diet for 4 weeks. RESULTS: Subacute toxicity, hematology, liver function, and organ weight tests in low-sodium sausage groups showed results similar to those of the control group, and all toxicity test levels were within normal ranges. CONCLUSIONS: All low-sodium sausage types tested are suggested to be safe in terms of subacute toxicity. Moreover, low-sodium sausages can be manufactured by biopolymer encapsulation of saltwort using pectin, chitosan, cellulose, and dextrin without toxicity. © 2019 Society of Chemical Industry.

Impact of partial substitution of NaCl by KCl, and MgCl2 on physicochemical and sensory properties of cooked sausages during storage.

OBJECTIVE: To determine the effect of three salting treatments (Control: 100% NaCl, T1: 60% NaCl, and 40% KCl, T2: 50% NaCl, 40% KCl, and 10% MgCl2) on meat quality of sausages during storage. METHODS: Various types of curing salts were used for processing of salamis. Physico-chemical and sensory evaluation of the sausages were analyzed during 4 weeks of ripening. RESULTS: The pH values and salinity contents were the highest in control, while they were the lowest in T2 at 4 weeks (p<0.05). Control and T1 had less thiobarbituric acid reactive substances (TBARS) than T2 at 2 weeks (p<0.05). The volatile basic nitrogen (VBN) levels in T1 were similar to control and lower than T2 at 2 and 4 weeks (p<0.05). Moreover, T1 samples were redder and had a higher saturation index (C*) value when compared to the others. T2 showed greater hardness, gumminess and chewiness when compared with the control (p<0.05). Control was comparable to T1 for sensory attributes. CONCLUSION: sodium chloride may be partially substituted with potassium chloride without negatively influencing meat quality of sausages up to 4 weeks of storage. These results could help to develop low-sodium sausages.

Controversy on the correlation of red and processed meat consumption with colorectal cancer risk: an Asian perspective.

This study aimed to investigate the relationship between meat intake and colorectal cancer risk from an Asian, particularly Korean, perspective. A report by the International Agency for Research on Cancer (IARC) published in 2015 concluded that intake of processed and red meat increases the risk of developing colorectal cancer. We conducted an in-depth analysis of prospective, retrospective, case-control and cohort studies, systematic review articles, and IARC monograph reports, which revealed that the IARC/WHO report weighted the results of studies based in Western countries more and that the correlation between intake of processed meat products and colorectal cancer incidence in Asians is not clearly supported. Among 73 epidemiological studies, approximately 76% were conducted in Western countries, whereas only 15% of studies were conducted in Asia. Furthermore, most studies conducted in Asia showed that processed meat consumption is not related to the onset of cancer. Moreover, there have been no reports showing significant correlation between various factors that directly or indirectly affect colorectal cancer incidence, including processed meat products types, raw meat types, or cooking methods. Further epidemiological studies taking each country's food culture into consideration are required to reliably elucidate the effects of processed meat product intake, especially on cancer incidence.

Mechanisms of Neuroprotective Effects of Peptides Derived from Natural Materials and Their Production and Assessment.

In this review, we provide an overview of the main mechanisms by which peptides derived from natural materials exhibit neuroprotective effects. We also review methods for the production of these peptides and various methods for determining their neuroprotective effects in vitro and in vivo. Neuroprotective peptides are closely associated with protection against apoptosis, with proteins related to the cell death/survival signaling pathway, with acetylcholine activity and with inhibition of oxidative stress and inflammation. Neuroprotective peptides derived from natural materials typically have a molecular weight below 1 kDa; they are most often derived using the extraction enzymes papain and alcalase. The neuroprotective peptides identified in the literature are mainly composed of hydrophobic amino acids with low molecular weight, with Asp and Glu at the N-terminal position. These observations may guide the development of novel peptides with potential neuroprotective effects.

Effect of Dietary Red Meat on Colorectal Cancer Risk-A Review.

Heme iron overload has been implicated as the main cause of the increased risk of cancer due to the consumption of red meat. However, fish and shellfish, teas, and spices contain up to five times more iron than red meat. There is insufficient evidence that iron intake in dietary red meat is the primary causal factor for colorectal cancer. In addition, harmful substances produced during the preparation of red meat, including heterocyclic amines (HCAs), polycyclic aromatic hydrocarbons (PAHs), N-nitroso compounds, and acrylamide, are extrinsic factors that increase carcinogenicity. HCAs are produced during the cooking of red meat, poultry meat, and fish. PAHs may also be produced during the cooking of diverse food groups, such as dairy products, fruits, vegetables, and cereals. The average daily intake of red meat among Korean individuals is 62 g; the amount of PAHs entering the body via red meat is less than the average amount of PAHs the body is exposed to in the air. Therefore, it is difficult to conclude that dietary red meat is the main cause of colorectal cancer. Rather, there may be an intricate influence of multiple factors, including fruit and vegetable intake, alcohol consumption, smoking, overweight, obesity, and stress.

Effects of Prunus mume Sieb. et Zucc. extract and its biopolymer encapsulation on a mouse model of colitis.

BACKGROUND: Prunus mume suppresses various diseases caused by inflammation response and exhibits antioxidant and free radical-scavenging activities. Therefore this study determined the effect of an aqueous P. mume (PM) extract in a mouse colitis model and investigated the value of biopolymer encapsulation, facilitating targeted delivery to the colon. Colitis was induced by administration of 30 g kg-1 dextran sulfate sodium to male BALB/c mice for 7 days prior to treatment with vehicle, 50 mg kg-1 PM extract or biopolymer-encapsulated PM extract, or 50 mg kg-1 sulfasalazine. RESULTS: Histological examination of the colon in BALB/c mice showed epithelial destruction and mucosal infiltration of inflammatory cells. These changes were attenuated in PM-treated mice, which had lower levels of inflammatory cytokines, cyclooxygenase 2 and immunoglobulins (IgA, IgM and IgE) compared with the vehicle-treated colitis group. The PM extract showed concentration-dependent radical scavenging and superoxide dismutase-like antioxidant activities. CONCLUSION: These results indicated that the effects of the PM extract on colitis were not influenced by biopolymer encapsulation and that this PM extract could be a potential therapeutic agent for inflammatory bowel disease. © 2016 Society of Chemical Industry.

Effects of self-carbon dioxide-generation material for active packaging on pH, water-holding capacity, meat color, lipid oxidation and microbial growth in beef during cold storage.

BACKGROUND: Active packaging refers to the mixing of additive agents into packaging materials with the purpose of maintaining or extending food product quality and shelf life. The aim of this study was to develop an easy and cheap active packaging for beef. Beef loin samples were divided into three packaging groups (C, ziplock bag packaging; T1, vacuum packaging; T2, active packaging) and stored at 4 °C for 21 days. RESULTS: The water-holding capacity was significantly (P < 0.05) higher in C and T2 than in T1 for up to 7 days of storage. The TBARS value was significantly (P < 0.05) lower in T1 and T2 after 7 days of storage. The counts of some microorganism were significantly (P < 0.05) lower in T1 and T2 after 7 days of storage; the total bacterial count and Escherichia coli count were lowest in T2 at the end of storage. CONCLUSION: These results indicate that active packaging using self-CO2 -generation materials can extend the shelf life similarly to that observed with vacuum packaging, and that the active packaging method can improve the quality characteristics of beef during cold storage. © 2017 Society of Chemical Industry.

Effect of the Ratio of Raw Material Components on the Physico-chemical Characteristics of Emulsion-type Pork Sausages.

This study was conducted to investigate the effects of raw material ratio on the physicochemical characteristics of emulsion-type pork sausages. Experiment design was divided into 12 treatments, based on protein level (P), fat level (3P, 3.5P, and 4P), and water level (4P+10, 4P+15, 4P+20, and 4P+25). The pH and shear force values were significantly higher in T7 (3.5P fat and 4P+20 water) than those of other treatments. The lightness and redness were greatly reduced by increasing the quantity of water. The treatments containing 3P fat and 4P+20 water had the highest values of cohesiveness, springiness, gumminess, and chewiness. On the whole, when the protein (P) and fat (3P, 3.5P, 4P) levels were fixed, an increase over the appropriate moisture level deteriorated many physicochemical characteristics.

Antioxidant, Liver Protective and Angiotensin I-converting Enzyme Inhibitory Activities of Old Laying Hen Hydrolysate in Crab Meat Analogue.

The purpose of this study was to evaluate the antioxidative activities of Crab meat analogue prepared with protein hydrolysates obtained from mechanically deboned chicken meat (MDCM) from spent laying hens. 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) radical-scavenging activity was increased by adding MDCM hydrolysates during storage, and activity correlated with the concentration of DPPH added up to 6 weeks of storage. Hydroxyl radical-scavenging activity was increased in all analogues containing MDCM hydrolysates. At 0 days of storage, angiotensin I-converting enzyme (ACE)-inhibitory activity was increased by the addition of MDCM hydrolysates. Activity did not correlate after 6 weeks of storage, in which ACE-inhibitory activity was increased with low concentrations of MDCM hydrolysates, but no ACE-inhibitory activity was observed at higher concentrations. The liver-protecting activity of crab meat analogue was shown to be around 60% of the positive control; however, it was not significantly different among the samples during storage. These results support the use of MDCM as a source of health-promoting constituents in crab meat analogue.

Low concentrations of doxycycline attenuates FasL-induced apoptosis in HeLa cells.

BACKGROUND: Doxycycline (DC) has been shown to possess non-antibiotic properties including Fas/Fas Ligand (FasL)-mediated apoptosis against several tumor types in the concentration range of 10-40 µg/mL. However, the effect of DC in apoptotic signaling at much low concentrations was not studied. METHODS: The present study investigated the attenuation effect of low dose of DC on FasL-induced apoptosis in HeLa cell by the methods of MTT assay, fluorescence microscopy, DNA fragmentation, flow cytometry analysis, and western blotting. RESULTS AND CONCLUSION: In the present findings we showed that low concentration of DC (<2.0 µg/mL) exhibited protective effects against FasL-induced apoptosis in HeLa cells. FasL treatment to HeLa cells resulted in a concentration-dependent induction of cell death, and treatment with low concentrations of DC (0.1-2 µg/mL) significantly (p < 0.001) attenuated the FasL-induced cell death as measured by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Further, the FasL-induced apoptotic features in HeLa cells, such as morphological changes, DNA fragmentation and cell cycle arrest was also inhibited by DC (0.5 µg/mL). Tetracycline and minocycline also showed similar anti-apoptotic effects but were not significant when compared to DC, tested at same concentrations. Further, DC (0.01-16 µg/mL) did not influence the hydrogen peroxide- or cisplatin-induced intrinsic apoptotic pathway in HeLa cells. Protein analysis using Western blotting confirmed that FasL-induced cleavage/activation of caspase-8 and caspase-3, were inhibited by DC treatment at low concentration (0.5 µg/mL). Considering the overall data, we report for the first time that DC exhibited anti-apoptotic effects at low concentrations in HeLa cells by inhibition of caspase activation via FasL-induced extrinsic pathway.

Current topics in active and intelligent food packaging for preservation of fresh foods.

The purpose of this review is to provide an overview of current packaging systems, e.g. active packaging and intelligent packaging, for various foods. Active packaging, such as modified atmosphere packaging (MAP), extends the shelf life of fresh produce, provides a high-quality product, reduces economic losses, including those caused by delay of ripening, and improves appearance. However, in active packaging, several variables must be considered, such as temperature control and different gas formulations with different product types and microorganisms. Active packaging refers to the incorporation of additive agents into packaging materials with the purpose of maintaining or extending food product quality and shelf life. Intelligent packaging is emerging as a potential advantage in food processing and is an especially useful tool for tracking product information and monitoring product conditions. Moreover, intelligent packaging facilitates data access and information exchange by altering conditions inside or outside the packaging and product. In spite of these advantages, few of these packaging systems are commercialized because of high cost, strict safety and hygiene regulations or limited consumer acceptance. Therefore more research is needed to develop cheaper, more easily applicable and effective packaging systems for various foods.

Impact of livestock industry on climate change: Case Study in South Korea - A review.

In recent years, there has been a growing argument attributing the primary cause of global climate change to livestock industry, which has led to the perception that the livestock industry is synonymous with greenhouse gas (GHG) emissions. However, a closer examination of the global GHG emission by sector reveals that the energy sector is responsible for the majority, accounting for 76.2% of the total, while agriculture contributes 11.9%. According to data from the Food and Agriculture Organization of the United Nations (FAO), the total GHG emissions associate with the livestock supply chain amount to 14.5%. Within this, emissions from direct sources, such as enteric fermentation and livestock manure treatment, which are not part of the front and rear industries, represent only 7%. Although it is true that the increase in meat consumption driven by global population growth and rising incomes, has contributed to higher methane (CH4) emissions resulting from enteric fermentation in ruminant animals, categorizing the livestock industry as the primary source of GHG emissions oversimplifies a complex issue and disregards objective data. Therefore, it may be a misleading to solely focus on the livestock sector without addressing the significant emissions from the energy sector, which is the largest contributor to GHG emissions. The top priority should be the objective and accurate measurement of GHG emissions, followed by the development and implementation of suitable reduction policies for each industrial sector with significant GHG emissions contributions.

Current Technologies and Future Perspective in Meat Analogs Made from Plant, Insect, and Mycoprotein Materials: A Review.

This study reviewed the current data presented in the literature on developing meat analogs using plant-, insect-, and protein-derived materials and presents a conclusion on future perspectives. As a result of this study, it was found that the current products developed using plant-, insect-, and mycoprotein-derived materials still did not provide the quality of traditional meat products. Plant-derived meat analogs have been shown to use soybean-derived materials and beta-glucan or gluten, while insect-derived materials have been studied by mixing them with plant-derived materials. It is reported that the development of meat analogs using mycoprotein is somewhat insufficient compared to other materials, and safety issues should also be considered. Growth in the meat analog market, which includes products made using plant-, insect-, and mycoprotein-derived materials is reliant upon further research being conducted, as well as increased efforts for it to coexist alongside the traditional livestock industry. Additionally, it will become necessary to clearly define legal standards for meat analogs, such as their classification, characteristics, and product-labeling methods.

Preliminary study on comparison of egg extraction methods for development of fetal bovine serum substitutes in cultured meat.

Fetal bovine serum (FBS) substitution remains one of the challenges to the realization of cultured meat production in the marketplace. In this study, three methods were developed to extract a substitute for FBS using egg white extract (EWE): using 25 mM CaCl2/2.5 % ammonium sulfate/citric acid (A); ethyl alcohol (B); and 5 % ammonium sulfate/citric acid (C). B EWE can effectively replace up to 50 % of FBS in growth media (10 % of the total). Ovalbumin in the extracts can promote cell proliferation, and components along the 12 kDa protein band have the potential to inhibit cell proliferation. Chick primary muscle cells applied with B EWE, an edible material that improved the cost and time efficiency of cultured meat production, effectively proliferated/differentiated. Therefore, EWE extracted using ethyl alcohol may be used as an FBS substitute to reduce animal sacrifices and should be considered a viable alternative to FBS for cultured meat.

Licochalcone A and B enhance muscle proliferation and differentiation by regulating Myostatin.

BACKGROUND: Myostatin (MSTN) inhibition has demonstrated promise for the treatment of diseases associated with muscle loss. In a previous study, we discovered that Glycyrrhiza uralensis (G. uralensis) crude water extract (CWE) inhibits MSTN expression while promoting myogenesis. Furthermore, three specific compounds of G. uralensis, namely liquiritigenin, tetrahydroxymethoxychalcone, and Licochalcone B (Lic B), were found to promote myoblast proliferation and differentiation, as well as accelerate the regeneration of injured muscle tissue. PURPOSE: The purpose of this study was to build on our previous findings on G. uralensis and demonstrate the potential of its two components, Licochalcone A (Lic A) and Lic B, in muscle mass regulation (by inhibiting MSTN), aging and muscle formation. METHODS: G. uralensis, Lic A, and Lic B were evaluated thoroughly using in silico, in vitro and in vivo approaches. In silico analyses included molecular docking, and dynamics simulations of these compounds with MSTN. Protein-protein docking was carried out for MSTN, as well as for the docked complex of MSTN-Lic with its receptor, activin type IIB receptor (ACVRIIB). Subsequent in vitro studies used C2C12 cell lines and primary mouse muscle stem cells to acess the cell proliferation and differentiation of normal and aged cells, levels of MSTN, Atrogin 1, and MuRF1, and plasma MSTN concentrations, employing techniques such as western blotting, immunohistochemistry, immunocytochemistry, cell proliferation and differentiation assays, and real-time RT-PCR. Furthermore, in vivo experiments using mouse models focused on measuring muscle fiber diameters. RESULTS: CWE of G. uralensis and two of its components, namely Lic A and B, promote myoblast proliferation and differentiation by inhibiting MSTN and reducing Atrogin1 and MuRF1 expressions and MSTN protein concentration in serum. In silico interaction analysis revealed that Lic A (binding energy -6.9 Kcal/mol) and B (binding energy -5.9 Kcal/mol) bind to MSTN and reduce binding between it and ACVRIIB, thereby inhibiting downstream signaling. The experimental analysis, which involved both in vitro and in vivo studies, demonstrated that the levels of MSTN, Atrogin 1, and MuRF1 were decreased when G. uralensis CWE, Lic A, or Lic B were administered into mice or treated in the mouse primary muscle satellite cells (MSCs) and C2C12 myoblasts. The diameters of muscle fibers increased in orally treated mice, and the differentiation and proliferation of C2C12 cells were enhanced. G. uralensis CWE, Lic A, and Lic B also promoted cell proliferation in aged cells, suggesting that they may have anti-muslce aging properties. They also reduced the expression and phosphorylation of SMAD2 and SMAD3 (MSTN downstream effectors), adding to the evidence that MSTN is inhibited. CONCLUSION: These findings suggest that CWE and its active constituents Lic A and Lic B have anti-mauscle aging potential. They also have the potential to be used as natural inhibitors of MSTN and as therapeutic options for disorders associated with muscle atrophy.

Targeting myostatin using quercetin as a media supplement to improve myogenesis for cultured meat production: An in silico and in vitro study.

Cultured meat (CM) is an alternative protein food and is produced by cultivating muscle satellite (stem) cells (MSCs) derived from livestock animals (bovine, chickens, and porcine) through myogenesis leading to generate muscle mass. Myostatin (MSTN) is well well-known negative regulator of myogenesis, and in the present study, in silico screening of natural compounds was performed to identify MSTN inhibitors. Interestingly, quercetin was found to inhibit MSTN (binding energy -7.40 kcal/mol), and this was further validated by a 100 ns molecular dynamics simulation. Quercetin was added to culture media to boost myogenesis, and its potent antioxidant property helped maintain media pH. Furthermore, quercetin increased the myotube thickness and length, increased MSC differentiation, and upregulated the gene and protein expressions of myoblast determination protein 1 (MYOD), Myogenin (MYOG), and Myosin heavy chains (MYH) in vitro. In addition, quercetin inhibited the activities of MSTN, activin receptor type-2B (ACVR2B), and SMAD2 and 3, and thus significantly enhanced MSC differentiation and myotube formation. Overall, this study shows that quercetin might be useful for enhancing large-scale CM production. It is hoped that this study provides a starting point for research in the CM area aimed to enhancing product quality, nutritional values, and the efficacy of large-scale production.

Current technology and industrialization status of cell-cultivated meat.

Interest and investment in cultivated meat are increasing because of the realization that it can effectively supply sufficient food resources and reduce the use of livestock. Nevertheless, accurate information on the specific technologies used for cultivated meat production and the characteristics of cultivated meat is lacking. Authorization for the use of cultivated meat is already underway in the United States, Singapore, and Israel, and other major countries are also expected to approve cultivated meat as food once the details of the intricate process of producing cultivated meat, which encompasses stages such as cell proliferation, differentiation, maturation, and assembly, is thoroughly established. The development and standardization of mass production processes and safety evaluations must precede the industrialization and use of cultivated meat as food. However, the technology for the industrialization of cultivated meat is still in its nascent stage, and the mass production process has not yet been established. The mass production process of cultivated meat may not be easy to disclose because it is related to the interests of several companies or research teams. However, the overall research flow shows that equipment development for mass production and cell acquisition, proliferation, and differentiation, as well as for three-dimensional production supports and bioreactors have not yet been completed. Therefore, additional research on the mass production process and safety of cultivated meat is essential. The consumer's trust in the cultivated meat products and production technologies recently disclosed by some companies should also be analyzed and considered for guiding future developments in this industry. Furthermore, close monitoring by academia and the government will be necessary to identify fraud in the cultivated meat industry.

Crusting-fabricated three-dimensional soy-based scaffolds for cultured meat production: A preliminary study.

Crusting has been developed as a non-chemical and non-machine intensive scaffold fabrication method. This method is based on the self-assembling ability of soy biomolecules, allowing the fabrication of a three-dimensional network for cell growth. Preliminary characterization revealed differences in pore size, water absorption, and degradation between pure soy-based scaffold (Y2R) and with added glycerol (Y2G). The Fourier-transform infrared spectrum absorbance peaks of functional groups related to proteins, carbohydrates, and lipids hinted the integration of soy biomolecules potentially via the Maillard reaction, as supported by the visible browning of the scaffold surface. Microscopic images revealed aligned myotubes in both scaffolds, with Y2G myotubes having greater proximity after 72 h of proliferation. Both spontaneous and electro-stimulated contractions were recorded as early as 72 h in proliferation medium. Crusting-fabricated soy-based scaffolds can further be explored for its application in cultured meat production.