Table 1. Summary of current status of technical issues and future required improvements for IMA

S.No.	Technical requirements	Current status	Required approaches
1	Cell bank	Except for Homo sapiens other primary cells are not reposited. No other primary muscle or adipose cell line is submitted.	Currently research for production is carried for bovine, porcine, chicken, fish, shrimps, horse, duck, kangaroo, rats. Central cell bank should be created and maintained for all the culture meat cell lines will be carried for research and production.
2	Database	No reference database is available except published research articles	Reference range for every IMA muscle models, Muscle cells, Adipose cells, Specifications for Proliferation, Differentiation, Required Biomaterials, Culture conditions, 2D and 3D culture requirements, material requirements, Contaminants including chemicals, physical and biological, Texture, taste, Nutritional value, Source of animals, genetic information and regulatory systems.
3	Culturing media	1) Currently used media are generalized media which suits every cell type but not specifically for muscle cells.2) No specific medium for proliferation or differentiation is available for different type of animal muscle or adipose.3) High cost	1) Animal type and cell type specific culture media should be engineered understanding the requirements based on genetic predisposition.2) Both proliferation and differentiation are different state of cell existence this the nutrition requirements, thus based on the cell metabolism and gene expression profile the required nutrition composition has to be formulated.3) Instead of animal source and plant source, using cloning technologies for growth factors or othr protein components for large scale will help in cost reduction.4) Co-culture supporting medium.
4	Scaffold & Microcarriers for 3D culture	1) Non-edible scaffolds or microcarriers are synthesized with edible materials due to the production methods.2) High cost for material and production.3) Labor intensive and still at research scale thus not readily available.	1) Edible and digestible scaffolds or microcarriers should be produced with food grade materials and methods.2) Produced materials should mimic the tissue or meat rheological properties.3) Stability of materials (temperature, sheer stress, shelf life).4) Animal-free materials are recommended.5) Nutrition and oxygen perfusion should be perfected.6) Mimic systemic vascularization.7) Water holding capacity.8) Less cooking loss.
5	Regulatory	No complete guidelines or regulations are framed for productions or for IMA research.	Complete guidelines and regulatory documentation should be made as to every aspect for raw materials, cell lines or bank, productions methods, media usage, etc.
6	Proliferation and differentiation	Primely based on the serum. concentration and very few factors like Insulin, EGF, transferrin, selenium, p38, etc., supplements.	1) Specific hub proteins to direct a progenitor or stem cell towards proliferative or differentiation state.2) Methods to direct the biochemical pathways for higher production.3) Inducers to fasten the cell cycle to change from progenitor state to proliferative state and differentiation state.This process is critical to control the cost and production issues.
7	Serum	1) Batch to batch variation,2) Supply and demand3) Ethical and animal welfare	1) Nutrition defined serum free medium for different cell state.2) Alternative to serum from non-animal source.3) Synthetic serum rather than animal source.
8	Anti-microbials	High proportions of anti-bacterial, anti-fungal and Antibiotics which are in general clinical use, as increased use of antibiotics will create drug resistance for clinical settings, inflated cost, cell stress.	Alternatives to antibiotics for example anti-microbial peptides, lysins, bacteriocins, SMAMP’s, IDR peptides, biological extracts, which will not be a stress factor or create drug resistance.
9	Bioreactors	1) Operational cost2) Sheer stress3) Nutrition diffusion4) Scalability5) Viability6) Quality7) labor intensive8) Contamination9) Optimization10) Storage11) Mobilization	Bioreactors issues are complex and bottle necks with various factors. Every stage should be carefully monitored and optimized. Any issue in the reactor leads to huge economic issues. Thus, every stage must be optimized for betterment and production of cells.