Abstract
One of the critical requirements for the production of artificial meat is to mimic the flavor of the original meat. Therefore, incorporation of heme has been proposed, but it is toxic when added at high concentrations in cell culture systems. Additionally, obtaining heme can be costly depending on the source. In this study, we aimed to support the growth of normal bovine muscle cells (BRMC-F2401, BRMCs) by introducing a bacterial extract obtained from a safe, high-heme-containing Corynebacterium species. The BRMCs exhibited heme toxicity when the bacterial heme level was >20 μM; however, they were adapted to stably proliferate with a 3 on-3 off culture scheme. RNA sequencing of the heme-adapted BRMCs showed gene expression changes, including upregulation of detoxification genes, CYP1A1, CYP26B1, and SERPINB2. The knockdown of these three genes increased heme sensitivity and reversed heme tolerance of the heme-adapted BRMCs. Additionally, ROS levels increased upon heme treatment, suggesting that ROS is an important factor in heme adaptation processes. Collectively, our study presents an affordable strategy to incorporate heme in cultured meat production and the mechanism underlying this process.