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首页> 外文期刊>Stem Cell Research & Therapy >Hepatocyte growth factor (HGF) and stem cell factor (SCF) maintained the stemness of human bone marrow mesenchymal stem cells (hBMSCs) during long-term expansion by preserving mitochondrial function via the PI3K/AKT, ERK1/2, and STAT3 signaling pathways
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Hepatocyte growth factor (HGF) and stem cell factor (SCF) maintained the stemness of human bone marrow mesenchymal stem cells (hBMSCs) during long-term expansion by preserving mitochondrial function via the PI3K/AKT, ERK1/2, and STAT3 signaling pathways

机译：肝细胞生长因子（HGF）和干细胞因子（SCF）通过通过PI3K / AKT，ERK1 / 2和Stat3信号传导途径保持线粒体函数，在长期膨胀期间保持人骨髓间充质干细胞（HBMSC）的茎。

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BACKGROUND:Mesenchymal stem cells (MSCs) have a limited self-renewal ability, impaired multi-differentiation potential, and undetermined cell senescence during in vitro series expansion. To address this concern, we investigated the effects of the microenvironment provided by stem cells from human exfoliated deciduous teeth (SHED) in maintaining the stemness of human bone marrow mesenchymal stem cells (hBMSCs) and identified the key factors and possible mechanisms responsible for maintaining the stemness of MSCs during long-term expansion in vitro.METHODS:The passage 3 (P3) to passage 8 (P8) hBMSCs were cultured in the conditioned medium from SHED (SHED-CM). The percentage of senescent cells was evaluated by β-galactosidase staining. In addition, the osteogenic differentiation potential was analyzed by reverse transcription quantitative PCR (RT-qPCR), Western blot, alizarin red, and alkaline phosphatase (ALP) staining. Furthermore, RT-qPCR results identified hepatocyte growth factor (HGF) and stem cell factor (SCF) as key factors. Thus, the effects of HGF and SCF on mitochondrial function were assessed by measuring the ROS and mitochondrial membrane potential levels. Finally, selected mitochondrial-related proteins associated with the PI3K/AKT, ERK1/2, and STAT3 signaling pathways were investigated to determine the effects of HGF and SCF in preserving the mitochondrial function of hBMSCs during long-term expansion.RESULTS:SHED-CM had significantly enhanced the cell proliferation, reduced the senescent cells, and maintained the osteogenesis and pro-angiogenic capacity in P8 hBMSCs during long-term expansion. In addition, hBMSCs treated with 100?ng/ml HGF and 10?ng/ml SCF had reduced ROS levels and preserved mitochondrial membrane potential compared with P8 hBMSCs during long-term expansion. Furthermore, HGF and SCF upregulated the expression of mitochondrial-related proteins associated with the PI3K/AKT, ERK1/2, and STAT3 signaling pathways, possibly contributing to the maintenance of hBMSCs stemness by preserving mitochondrial function.CONCLUSION:Both HGF and SCF are key factors in maintaining the stemness of hBMSCs by preserving mitochondrial function through the expression of proteins associated with the PI3K/AKT, ERK1/2, and STAT3 signaling pathways. This study provides new insights into the anti-senescence capability of HGF and SCF, as well as new evidence for their potential application in optimizing the long-term culture of MSCs.
机译：背景：间充质干细胞（MSCs）具有有限的自我更新能力，多分化潜力受损，并且在体外串联膨胀期间衰老未确定的细胞衰老。为了解决这种问题，我们研究了干细胞提供的微环境从人脱色落叶齿（SHED）的效果保持人骨髓间充质干细胞（HBMSCs）的茎，并确定了负责维护的关键因素和可能机制在体外长期膨胀期间MSCs的茎。方法：将通道3（P3）与通道8（P8）HBMSCs在来自Shed（Shed-cm）的条件培养基中培养。通过β-半乳糖苷酶染色评价衰老细胞的百分比。此外，通过逆转录定量PCR（RT-QPCR），Western印迹，茜素红色和碱性磷酸酶（ALP）染色来分析骨质发生分化潜力。此外，RT-QPCR结果将肝细胞生长因子（HGF）和干细胞因子（SCF）确定为关键因素。因此，通过测量ROS和线粒体膜电位水平来评估HGF和SCF对线粒体功能的影响。最后，研究了与PI3K / AKT，ERK1 / 2和STAT3信号传导途径相关的选定的线粒体相关蛋白，以确定HGF和SCF在长期扩展期间保持HBMSCS的线粒体功能的影响。结果：棚屋已经显着增强了细胞增殖，降低了衰老细胞，并在长期膨胀期间保持了P8 HBMSCS中的骨开发和促血管生成能力。另外，用100·Ng / ml HGF和10·Ng / ml SCF处理的HBMSCs具有降低的ROS水平，并在长期膨胀期间与P8 HBMSCS相比保存了线粒体膜电位。此外，HGF和SCF上调了与PI3K / AKT，ERK1 / 2和STAT3信号传导途径相关的线粒体相关蛋白的表达，可能通过保留线粒体函数来维持HBMSCs茎的维持。结论：HGF和SCF都是关键通过将线粒体通过与PI3K / AKT，ERK1 / 2和Stat3信号传导途径相关的蛋白质保持线粒体函数来维持HBMSCs茎的因素。本研究提供了新的洞察HGF和SCF的抗衰老能力，以及在优化MSC的长期文化方面潜在应用的新证据。

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