Schive Simen W, Mirlashari Mohammad Reza, Hasvold Grete, Wang Mengyu, Josefsen Dag, Gullestad Hans Petter, Korsgren Olle, Foss Aksel, Kvalheim Gunnar, Scholz Hanne
Department of Transplant Medicine, Oslo University Hospital, Oslo, Norway.
†Institute for Surgical Research, Oslo University Hospital, Oslo, Norway.
Cell Med. 2017 Apr 14;9(3):103-116. doi: 10.3727/215517917X693401. eCollection 2017.
Adipose-derived mesenchymal stem cells (ASCs) release factors beneficial for islets in vitro and protect against hyperglycemia in rodent models of diabetes. Oxygen tension has been shown to induce metabolic changes and alter ASCs' release of soluble factors. The effects of hypoxia on the antidiabetic properties of ASCs have not been explored. To investigate this, we incubated human ASCs for 48 h in 21% (normoxia) or 1% O (hypoxia) and compared viability, cell growth, surface markers, differentiation capability, and soluble factors in the conditioned media (CM). Human islets were exposed to CM from ASCs incubated in either normoxia or hypoxia, and islet function and apoptosis after culture with or without proinflammatory cytokines were measured. To test hypoxic preconditioned ASCs' islet protective effects in vivo, ASCs were incubated for 48 h in normoxia or hypoxia before being injected into Balb/c Rag 1 immunodeficient mice with streptozotocin-induced insulitis. Progression of diabetes and insulin content of pancreas were measured. We found that incubation in hypoxia was well tolerated by ASCs and that levels of VEGF-A, FGF-2, and bNGF were elevated in CM from ASCs incubated in hypoxia compared to normoxia, while levels of HGF, IL-8, and CXCL1 were reduced. CM from ASCs incubated in hypoxia significantly improved human islet function and reduced apoptosis after culture, and reduced cytokine-induced apoptosis. In our mouse model, pancreas insulin content was higher in both groups receiving ASCs compared to control, but the mice receiving preconditioned ASCs had lower random and fasting blood glucose, as well as improved oral glucose tolerance compared to untreated mice. In conclusion, our in vitro results indicate that the islet protective potential of ASCs improves in hypoxia, and we give insight into factors involved in this. Finally we show that hypoxic preconditioning potentiates ASCs' antidiabetic effect in vivo.
脂肪来源的间充质干细胞（ASCs）在体外释放对胰岛有益的因子，并在糖尿病啮齿动物模型中预防高血糖。氧张力已被证明可诱导代谢变化并改变ASCs可溶性因子的释放。低氧对ASCs抗糖尿病特性的影响尚未得到研究。为了探究这一点，我们将人ASCs在21%（常氧）或1% O₂（低氧）条件下孵育48小时，并比较了其活力、细胞生长、表面标志物、分化能力以及条件培养基（CM）中的可溶性因子。将人胰岛暴露于常氧或低氧孵育的ASCs的CM中，并测量在有或无促炎细胞因子培养后的胰岛功能和凋亡情况。为了测试低氧预处理的ASCs在体内的胰岛保护作用，将ASCs在常氧或低氧条件下孵育48小时，然后注射到链脲佐菌素诱导的胰岛炎的Balb/c Rag 1免疫缺陷小鼠体内。测量糖尿病的进展和胰腺的胰岛素含量。我们发现低氧孵育对ASCs耐受性良好，与常氧相比，低氧孵育的ASCs的CM中VEGF-A、FGF-2和bNGF水平升高，而HGF、IL-8和CXCL1水平降低。低氧孵育的ASCs的CM显著改善了人胰岛功能，并减少了培养后的凋亡，以及减少了细胞因子诱导的凋亡。在我们的小鼠模型中，与对照组相比，接受ASCs的两组胰腺胰岛素含量均较高，但与未处理的小鼠相比，接受预处理ASCs的小鼠随机和空腹血糖较低，口服葡萄糖耐量也有所改善。总之，我们的体外结果表明，低氧条件下ASCs的胰岛保护潜力增强，我们深入了解了其中涉及的因素。最后，我们表明低氧预处理增强了ASCs在体内的抗糖尿病作用。
