Tejedor Gautier, Luz-Crawford Patricia, Barthelaix Audrey, Toupet Karine, Roudières Sébastien, Autelitano François, Jorgensen Christian, Djouad Farida
IRMB, INSERM, University of Montpellier, Montpellier, France.
Laboratorio de Inmunología Celular y Molecular, Facultad de Medicina, Universidad de los Andes, Santiago, Chile.
Front Cell Dev Biol. 2021 Mar 2;9:579951. doi: 10.3389/fcell.2021.579951. eCollection 2021.
The super healer Murphy Roths Large (MRL) mouse represents the "holy grail" of mammalian regenerative model to decipher the key mechanisms that underlies regeneration in mammals. At a time when mesenchymal stem cell (MSC)-based therapy represents the most promising approach to treat degenerative diseases such as osteoarthritis (OA), identification of key factors responsible for the regenerative potential of MSC derived from MRL mouse would be a major step forward for regenerative medicine. In the present study, we assessed and compared MSC derived from MRL (MRL MSC) and C57BL/6 (BL6 MSC) mice. First, we compare the phenotype and the differentiation potential of MRL and BL6 MSC and did not observe any difference. Then, we evaluated the proliferation and migration potential of the cells and found that while MRL MSC proliferate at a slower rate than BL6 MSC, they migrate at a significantly higher rate. This higher migration potential is mediated, in part, by MRL MSC-secreted products since MRL MSC conditioned medium that contains a complex of released factors significantly increased the migration potential of BL6 MSC. A comparative analysis of the secretome by quantitative shotgun proteomics and Western blotting revealed that MRL MSC produce and release higher levels of mesencephalic astrocyte-derived neurotrophic factor (MANF) as compared to MSC derived from BL6, BALB/c, and DBA1 mice. MANF knockdown in MRL MSC using a specific small interfering RNA (siRNA) reduced both MRL MSC migration potential in scratch wound assay and their regenerative potential in the ear punch model in BL6 mice. Finally, injection of MRL MSC silenced for MANF did not protect mice from OA development. In conclusion, our results evidence that the enhanced regenerative potential and protection from OA of MRL mice might be, in part, attributed to their MSC, an effective reservoir of MANF.
超级愈合型墨菲罗斯大（MRL）小鼠代表了哺乳动物再生模型中的“圣杯”，有助于破解哺乳动物再生的关键机制。在基于间充质干细胞（MSC）的疗法成为治疗骨关节炎（OA）等退行性疾病最具前景的方法之际，鉴定源自MRL小鼠的MSC的再生潜能的关键因素将是再生医学向前迈出的重要一步。在本研究中，我们评估并比较了源自MRL（MRL MSC）和C57BL/6（BL6 MSC）小鼠的MSC。首先，我们比较了MRL和BL6 MSC的表型及分化潜能，未观察到任何差异。然后，我们评估了细胞的增殖和迁移潜能，发现虽然MRL MSC的增殖速度比BL6 MSC慢，但它们的迁移速度明显更高。这种更高的迁移潜能部分是由MRL MSC分泌的产物介导的，因为含有释放因子复合物的MRL MSC条件培养基显著提高了BL6 MSC的迁移潜能。通过定量鸟枪法蛋白质组学和蛋白质印迹对分泌组进行的比较分析显示，与源自BL6、BALB/c和DBA1小鼠的MSC相比，MRL MSC产生和释放更高水平的中脑星形胶质细胞源性神经营养因子（MANF）。使用特异性小干扰RNA（siRNA）敲低MRL MSC中的MANF，在划痕试验中降低了MRL MSC的迁移潜能，在BL6小鼠的耳打孔模型中降低了它们的再生潜能。最后，注射沉默了MANF的MRL MSC并不能保护小鼠免受OA的发展。总之，我们的结果证明，MRL小鼠增强的再生潜能和对OA的保护作用可能部分归因于它们的MSC，即MANF的有效储存库。
