College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
College of Food Science and Technology, Nanjing Agricultural University National Center of Meat Quality and Safety Nanjing, MOST Key Laboratory of Meat Processing and Quality Control, MOE Key Laboratory of Meat Processing, MOA Nanjing 210095, China.
Food Res Int. 2023 Apr;166:112606. doi: 10.1016/j.foodres.2023.112606. Epub 2023 Feb 16.
Cultured meat is an efficient, safe and sustainable meat production technology. Adipose-derived stem cell (ADSC) is a promising cell type for cultured meat. In vitro, obtaining numerous of ADSCs is a pivotal step for cultured meat. In this research, we demonstrated that the proliferation and adipogenic differentiation of ADSCs significantly decreased during serial passage. Then, senescence β-galactosidase (SA-β-gal) staining showed that the positive rate of P9 ADSCs was 7.74-fold than P3 ADSCs. Subsequently, RNA sequencing (RNA-seq) was performed for P3 and P9 ADSCs and found that PI3K-AKT pathway was up-regulated, but cell cycle and DNA repair pathway were down-regulated in P9 ADSCs. Then, N-Acetylcysteine (NAC) was added during long-term expansion and showed that NAC enhanced the ADSCs proliferation and maintained adipogenic differentiation. Finally, RNA-seq was performed for P9 ADSCs cultured with or without NAC and showed that NAC restored the cell cycle and DNA repair pathway in P9 ADSCs. These results highlighted that NAC was an excellent supplement for large-scale expansion of porcine ADSCs for cultured meat.
培养肉是一种高效、安全且可持续的肉类生产技术。脂肪来源的干细胞（ADSC）是培养肉有前途的细胞类型。在体外，获得大量 ADSC 是培养肉的关键步骤。在这项研究中，我们证明 ADSC 的增殖和脂肪分化能力在传代过程中显著下降。然后，衰老β-半乳糖苷酶（SA-β-gal）染色显示 P9 ADSC 的阳性率是 P3 ADSC 的 7.74 倍。随后，对 P3 和 P9 ADSC 进行 RNA 测序（RNA-seq），发现 PI3K-AKT 通路在 P9 ADSC 中上调，而细胞周期和 DNA 修复通路在 P9 ADSC 中下调。然后，在长期扩增过程中添加 N-乙酰半胱氨酸（NAC），结果表明 NAC 增强了 ADSC 的增殖能力并维持了其脂肪分化能力。最后，对用或不用 NAC 培养的 P9 ADSC 进行 RNA-seq 分析，结果表明 NAC 恢复了 P9 ADSC 中的细胞周期和 DNA 修复通路。这些结果表明，NAC 是大规模扩增用于培养肉的猪 ADSC 的理想补充剂。
