胎牛血清(FBS),含有各种营养素,包含20%的细胞培养肉生长培养基。然而,伦理,成本,和科学问题,需要识别替代品。在这项研究中,我们研究了能够培养Hanwoo卫星细胞(HWSC)的商业制造的无血清培养基,以鉴定成分增殖增强因子.选择六种不同的无血清培养基,并将这些无血清培养基中的HWSC增殖速率与补充有20%FBS的对照培养基进行比较。在六家媒体中,仅在StemFlexTM培养基(SF)和间充质干细胞生长培养基DXF(MS)中的细胞增殖率高于对照培养基。SF和MS含有高成纤维细胞生长因子2(FGF2)浓度,我们发现在SF或MS中培养的细胞中FGF2蛋白表达上调。成纤维细胞生长因子受体1(FGFR1)介导的信号通路的激活和肌肉卫星细胞增殖相关因子的刺激被相关生物标志物的存在证实(FGFR1,FRS2,Raf1,ERK,p38、Pax7和MyoD),如定量聚合酶链反应所示,西方印迹,和免疫细胞化学.此外,PD173074,一种FGFR1抑制剂抑制了SF和MS中的细胞增殖,并下调了相关的生物标志物(FGFR1,FRS2,Raf1和ERK)。因此,SF和MS中细胞增殖的促进归因于FGF2,这表明肌肉卫星细胞中的FGFR1活化可能是提高细胞培养肉生产效率的目标。
Fetal bovine serum (FBS), which contains various nutrients, comprises 20% of the growth medium for cell-cultivated meat. However, ethical, cost, and scientific issues, necesitates identification of alternatives. In this study, we investigated commercially manufactured serum-free media capable of culturing Hanwoo satellite cells (HWSCs) to identify constituent proliferation enhancing factors. Six different serum-free media were selected, and the HWSC proliferation rates in these serum-free media were compared with that of control medium supplemented with 20% FBS. Among the six media, cell proliferation rates were higher only in StemFlexTM Medium (SF) and Mesenchymal Stem Cell Growth Medium DXF (MS) than in the control medium. SF and MS contain high fibroblast growth factor 2 (FGF2) concentrations, and we found upregulated FGF2 protein expression in cells cultured in SF or MS. Activation of the fibroblast growth factor receptor 1 (FGFR1)-mediated signaling pathway and stimulation of muscle satellite cell proliferation-related factors were confirmed by the presence of related biomarkers (FGFR1, FRS2, Raf1, ERK, p38, Pax7, and MyoD) as indicated by quantitative polymerase chain reaction, western blotting, and immunocytochemistry. Moreover, PD173074, an FGFR1 inhibitor suppressed cell proliferation in SF and MS and downregulated related biomarkers (FGFR1, FRS2, Raf1, and ERK). The promotion of cell proliferation in SF and MS was therefore attributed to FGF2, which indicates that FGFR1 activation in muscle satellite cells may be a target for improving the efficiency of cell-cultivated meat production.