延长体外复制寿命并保护分化和分泌能力，这是来自美国 Department of Pathology, The Ohio State University, Columbus, Ohio 43210。这样美国就已经有四个大学开展氢气效应的研究。

哈佛大学（2）、匹兹堡大学（5）、罗马林达大学（2）和俄亥俄州立大学（1）。现在已经有10篇论文，不过美国在论文数量上潜力非常大，因为是多个比较厉害的学校先后开展这个工作。他们一般都习惯延续性工作。

从研究特点上看，美国学者研究的比较细致，推理比较严谨。值得学习

使用骨髓间充质干细胞进行细胞治疗是再生医学的重要手段。骨髓间充质干细胞数量有限，需要体外增殖才能使用，但是这种操作可导致细胞老化，造成分化、增殖和治疗作用的丢失。氢气分子具有器官保护作用，该作用与氢气可选择性清除羟基自由基有关。氧化应激是导致细胞在体和离体老化的关键因素。本研究假定氢气能保护细胞体外增殖过程的老化。通过在培养系统中（气体介质）增加3%氢气的办法，发现可在对细胞培养早期克隆形成和复制寿命均有改善作用，同时能保护细胞分化和旁分泌能力。不过有意思的是，研究发现氢气并没有减低羟基自由基，对蛋白和核酸氧化损伤指标也没有影响，提示氢气对骨髓间充质干细胞的上述作用与其抗氧化作用无关。（对干细胞的有关翻译不一定准确，欢迎拍）

    这个文章提供了一种氢气效应细胞学研究手段，含3%氢气的混合气持续培养的方法，这与日本学者最早研究的细胞学方法明显不同，也更人性化，比较容易实现，但要注意的是两者的氢气浓度相差十分大，这个3%在日本学者的研究中应该属于没有效果的浓度，但这个研究是长时间培养的。

   因此，从这个角度考虑，氢气的作用与浓度、作用时间有非常大的关联性，这就更说明氢气是一种十分强大的生物活性分子。更值得称道的是，这个研究没有看到氢气的抗氧化作用，这又给我们的研究带来了更多遐想的空间。不是抗氧化，又是什么机制？

2.2. Hydrogen gas treatment

Hydrogen gas treatment was made by culturing cells in premixed gas (3% H₂, 21% O₂, 5% CO₂, balance N₂) (Praxair, Danbury, CT) in a hypoxia chamber (Stemcell Technologies, Vancouver, Canada). In brief, cell culture dishes or multiwell plates were placed in the chamber equipped with airtight seal. Then, the chamber was flushed for 5 min or more with premixed gas (20 L/min) according to the manufacture’s instruction. The chamber was flushed with premixed gas every 3 days or less to ensure the composition of mixed gas.

 

 

 

Hydrogen gas treatment prolongs replicative lifespan of bone marrow multipotential stromal cells in vitro while preserving differentiation and paracrine potentials
Biochemical and Biophysical Research Communications, In Press, Accepted Manuscript, Available online 4 June 2010
Haruhisa Kawasaki, Jianjun Guan, Kenichi Tamama

 Department of Pathology, The Ohio State University, Columbus, Ohio 43210

Cell therapy with bone marrow multipotential stromal cells / mesenchymal stem cells (MSCs) represents a promising approach in the field of regenerative medicine. Low frequency of MSCs in adult bone marrow necessitates ex vivo expansion of MSCs after harvest; however, such a manipulation causes cellular senescence with loss of differentiation, proliferative, and therapeutic potentials of MSCs. Hydrogen molecules have been shown to exert organ protective effects through selective reduction of hydroxyl radicals. As oxidative stress is one of the key insults promoting cell senescence in vivo as well as in vitro, we hypothesized that hydrogen molecules prevent senescent process during MSC expansion. Addition of 3%hydrogen gas enhanced preservation of colony forming early progenitor cells within MSC preparation and prolonged the in vitro replicative lifespan of MSCs without losing differentiation potentials and paracrine capabilities. Interestingly, 3%hydrogen gas treatment did not decrease hydroxyl radical, protein carbonyl, and 8-hydroxydeoxyguanosine, suggesting that scavenging hydroxyl radical might not be responsible for these effects of hydrogen gas in this study.

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