LabMeatNow’s Animal-Free Media for Culturing Chicken Skeletal Muscle Cells

The usual growth media for cell cultures contains 

• Dulbecco’s Modified Eagle Medium (DMEM): cell culture medium, provides essential nutrients and vitamins necessary for the growth
• Fetal bovine serum (FBS): serum supplement (growth factors, nutrients) for cell proliferation and differentiation.
• Penicillin-streptomycin: This is an antibiotic mixture 
• Insulin: hormone for cell growth and differentiation.
• Epidermal growth factor (EGF): growth factor for proliferation of muscle precursor cells.
• Hepatocyte growth factor (HGF): growth factor for muscle cell proliferation and differentiation.
• Glutamine: amino acid for cell growth and metabolism.

In summary, growth media consists of nutrients, salts, amino acids, vitamins, and growth factors.

Since growing meat from fetal bovine serum to replace conventional meat is unreasonable, lets look at an example of a fetal bovine serum-free media composition that has been used for culturing chicken skeletal muscle cells:

Basal media: DMEM/F-12 (1:1) or DMEM/high glucose

Supplements:

1. Insulin-transferrin-selenium (ITS) supplement (usually from FBS) replace with recombinant insulin
2. Bovine serum albumin 
3. L-carnitine
4. Creatine
5. Growth factors:
   • Epidermal growth factor (EGF)
   • Basic fibroblast growth factor (bFGF)
   • Hepatocyte growth factor (HGF)
   • Insulin-like growth factor-1 (IGF-1)
   • Platelet-derived growth factor (PDGF)

In the next step we want to avoid all animal derived products and improve the recipe:

Basal media: DMEM/F-12 (1:1) (or DMEM/high glucose)

Supplements:

1. Recombinant Insulin-Transferrin-Selenium
2. Glutamine 
3. Recombinant Growth factors (proteins):
   • Recombinant human EGF (Promotes cell proliferation and survival.)
   • Recombinant human bFGF (Supports proliferation of muscle progenitor cells.)
   • Recombinant human IGF-1 (Encourages muscle cell differentiation and growth.)
   • Recombinant human HGF (Helps in muscle regeneration and differentiation.)
   • Recombinant human PDGF (Stimulates proliferation and wound healing.)

Secondary Components (Enhance Growth and Differentiation)

1. Plant-based B27 supplement (contains a mixture of antioxidants, vitamins, hormones, and trace elements/minerals(separate!))
2. Plant-based N2 supplement (contains a mixture of hormones and trace elements)
3. Essential fatty acids linoleic acid + oleic acid + synthetic Cholesterol+ Polysorbate 80 as emulsifier
4. Ascorbic Acid (synthetic Vitamin C)
5. plant-based L-carnitine
6. plant-based Creatine (creature for example)
7. dexamethasone (for cells to mature into functional muscle tissue) 

Additionally we need to use silk-tissue as base matrix for the cells to adhere to, coated in adhesion proteins recombinantcollagen, recombinantfibronectin and recombinantlaminin (and sterilised by UV light)

In a next article we will look at each of these ingredients, identify which ones are too expensive or complicated to source, and replace those with simpler alternatives.

This I will do on another day. 

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