培养肉的成本有效开发的关键因素是可在无血清条件下培养的细胞系,以降低生产成本。培养肉中的血红素补充剂模仿原始肉的风味和颜色。这项研究引入了从棒状杆菌产生的细菌提取物,该细菌提取物通过定向进化被选择用于高血红素表达。正常的猪细胞系,PK15用于施用细菌血红素提取物作为补充剂。与先前的研究一致,我们观察到PK15对血红素提取物在10mM或更高的细胞毒性。然而,长期暴露后,PK15适于耐受高达40mM的血红素。对这些血红素适应的PK15细胞(PK15H)的RNA-seq分析揭示了一组改变的基因,主要参与细胞增殖,新陈代谢,和炎症。我们发现细胞色素P450家族1亚家族A,多肽1(CYP1A1),乳过氧化物酶(LPO),谷胱甘肽过氧化物酶5(GPX5)在PK15H血红素剂量依赖性上调。当我们将血清从2%连续降低到无血清时,我们得出了用5-10mM血红素提取物暂时维持的PK15H亚群。总之,我们的研究报告了一种可在高血红素培养基中培养的猪细胞,该细胞可在无血清条件下维持,并提出了在这一适应过程中起关键作用的标记基因.
A key element for the cost-effective development of cultured meat is a cell line culturable in serum-free conditions to reduce production costs. Heme supplementation in cultured meat mimics the original meat flavor and color. This study introduced a bacterial extract generated from Corynebacterium that was selected for high-heme expression by directed evolution. A normal porcine cell line, PK15, was used to apply the bacterial heme extract as a supplement. Consistent with prior research, we observed the cytotoxicity of PK15 to the heme extract at 10 mM or higher. However, after long-term exposure, PK15 adapted to tolerate up to 40 mM of heme. An RNA-seq analysis of these heme-adapted PK15 cells (PK15H) revealed a set of altered genes, mainly involved in cell proliferation, metabolism, and inflammation. We found that cytochrome P450, family 1, subfamily A, polypeptide 1 (CYP1A1), lactoperoxidase (LPO), and glutathione peroxidase 5 (GPX5) were upregulated in the PK15H heme dose dependently. When we reduced serum serially from 2% to serum free, we derived the PK15H subpopulation that was transiently maintained with 5-10 mM heme extract. Altogether, our study reports a porcine cell culturable in high-heme media that can be maintained in serum-free conditions and proposes a marker gene that plays a critical role in this adaptation process.