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Vitamin B12, iron, and other essential nutrients might be supplied by gut-colonizing bacteria.121 The in vitro meat product must also have an internal circulation medium to provide oxygen, nutrients, and waste products. E.62 is a standard medium that has been used for the growth and development of muscle tissue. Grown muscle tissue is believed to contain a few percent of fat cells.121 The amount of fat is believed to be energy-dense and a means of retaining nutrients.122 Besides, glucose, insulin, and other energy and growth regulators in muscle tissue also play an important role in the internal circulation medium. In fact, as human muscle tissue requires about 1,300 calories daily, the in vitro meat product must also be able to provide other nutrients to create an energy balance state for the muscle tissue cells. In vitro meat product must be a replacer of meat nutrients. Nutrients, such as energy (calories), protein, fiber, and other fatty acids, are the main forms of nutrition for muscle.123 Muscle tissue contains high levels of proteins, fat, and carbohydrates; thus, the energy source is very low in the first stages of cell development. The minimum requirement for energy is about 30 kcal per day, but for the final development of myotubes, the 143 kcal daily requirement for each quof myotubes should be added to allow for energy maintenance.124 The 20 kcal daily nutrition requirement of the cells can be obtained from protein-rich carbohydrate containing medium, which is lower in calories than animal protein.125 Thus, a protein-rich low-calorie medium is needed for myotubes to achieve the energy balance.126 By balance, we mean that the amount of nutrients being used by the cells should be equal to that being produced by the cells at a given time.127 Nutrients such as protein, carboxylic acids, and fatty acid esters are the main constituents of the muscle tissue.128
Gut microbiota is involved in the metabolism of cholesterol and in the synthesis of short-chain fatty acids (such as acetate, butyrate, and propionate). Thus, metabolites such as acetate and butyrate will provide energy and support for myotubes. d2c66b5586