Production of cultured meat from pig muscle stem cells

Haozhe Zhu; Zhongyuan Wu; Xi Ding; Mark J Post; Renpeng Guo; Jie Wang; Junjun Wu; Wenlai Tang; Shijie Ding; Guanghong Zhou

doi:10.1016/j.biomaterials.2022.121650

Production of cultured meat from pig muscle stem cells

Biomaterials. 2022 Aug:287:121650. doi: 10.1016/j.biomaterials.2022.121650. Epub 2022 Jul 18.

Authors

Haozhe Zhu¹, Zhongyuan Wu¹, Xi Ding¹, Mark J Post², Renpeng Guo³, Jie Wang⁴, Junjun Wu³, Wenlai Tang⁵, Shijie Ding⁶, Guanghong Zhou⁷

Affiliations

¹ College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China; National Center of Meat Quality and Safety Control, MOST; Key Laboratory of Meat Processing and Quality Control, MOE; Key Laboratory of Meat Processing, MOA, Nanjing Agricultural University, 210095, Jiangsu, China.
² Department of Physiology, Maastricht University, CARIM, Maastricht, the Netherlands.
³ College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China.
⁴ College of Artificial Intelligence, Nanjing Agricultural University, Nanjing, 210031, China.
⁵ School of Electrical and Automation Engineering, and Jiangsu Key Laboratory of 3D Printing Equipment and Manufacturing, Nanjing Normal University, Nanjing, 210023, China; State Key Laboratory of Pharmaceutical Biotechnology, Department of Sports Medicine and Adult Reconstructive Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, 210008, China. Electronic address: wltang@njnu.edu.cn.
⁶ College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China; National Center of Meat Quality and Safety Control, MOST; Key Laboratory of Meat Processing and Quality Control, MOE; Key Laboratory of Meat Processing, MOA, Nanjing Agricultural University, 210095, Jiangsu, China. Electronic address: shijieding@njau.edu.cn.
⁷ College of Food Science and Technology, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China; National Center of Meat Quality and Safety Control, MOST; Key Laboratory of Meat Processing and Quality Control, MOE; Key Laboratory of Meat Processing, MOA, Nanjing Agricultural University, 210095, Jiangsu, China. Electronic address: guanghong.zhou@hotmail.com.

PMID: 35872554
DOI: 10.1016/j.biomaterials.2022.121650

Abstract

Cultured meat is meat for consumption produced in a more sustainable way. It involves cell harvesting and expansion, differentiation into myotubes, construction into muscle fibres and meat structuring. We isolated 5.3 × 10⁴ porcine muscle stem cells from 1 g of neonatal pig muscle tissue. According to calculations, we need to expand muscle stem cells 10⁶-10⁷ times to produce 100 g or 1 kg of cultured meat. However, the cells gradually lost the ability to express stemness and mature muscle cell markers (PAX7, MyHC). To tackle this critical issue and maintain cell function during cell expansion, we found that long-term culture with (100 μM) l-Ascorbic acid 2-phosphate (Asc-2P) accelerated cell proliferation while preserving the muscle cell differentiation. We further optimized a scalable PDMS mold. Porcine muscle stem cells formed structurally-organized myotubes similar to muscle fibres in the mold. Asc-2P enhanced porcine muscle cells grown as 3D tissue networks that can produce a relatively large 3D tissue networks as cultured meat building blocks, which showed improved texture and amino acid content. These results established a realistic workflow for the production of cultured meat that mimics the pork meat structurally and is potentially scalable for industry.

Keywords: 3D tissue networks; Cultured meat; Porcine muscle satellite cells; Reactive oxygen species; l-ascorbic acid 2-phosphate.