Stem Cell Reviews and Reports ( IF 4.5 ) Pub Date : 2021-01-03 , DOI: 10.1007/s12015-020-10106-6 Robi Ar Bolli 1 , Chandrashekhar Dasari 1 , Asma Arshia 1 , Dinesh Devadoss 2 , Yiru Guo 1 , Usman Ashraf 1 , Qianhong Li 1
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Cardiac mesenchymal cells (CMCs), a newly-discovered and promising type of progenitor cells, are effective in improving cardiac function in rodents after myocardial infarction. Stem/progenitor cells are usually cultured at atmospheric O2 tension (21%); however, the physiologic O2 tension in the heart is ~5%, raising the concern that 21% O2 may cause toxicity due to oxidative stress. Thus, we compared mouse CMCs cultured at 21% or 5% O2 beginning at passage 2. At passage 5, CMCs underwent severe hypoxic stress (1% O2 for 24 h). Compared with CMCs cultured at 21% O2, culture at 5% O2 consistently improved cell morphology throughout 5 passages, markedly decreased cell size, increased cell number, shortened cell doubling time, and dramatically reduced lactate dehydrogenase release from CMCs into culture media after hypoxic stress. Furthermore, culture at 5% O2 increased telomerase activity and telomere length, implying that 21% O2 tension impairs telomerase activity, resulting in telomere shortening and decreased cell proliferation. Thus far, almost all preclinical and clinical studies of cell therapy for the heart disease have used atmospheric (21%) O2 to culture cells. Our data challenge this paradigm. Our results demonstrate that, compared with 21% O2, 5% O2 tension greatly enhances the competence and functional properties of CMCs. The increased proliferation rate at 5% O2 means that target numbers of CMCs can be achieved with much less time and cost. Furthermore, since this increased proliferation may continue in vivo after CMC transplantation, and since cells grown at 5% O2 are markedly resistant to severe hypoxic stress, and thus may be better able to survive after transplantation into scarred regions of the heart where O2 is very low, culture at 5% O2 may enhance the reparative properties of CMCs (and possibly other cell types). In conclusion, our data support a change in the methods used to culture CMCs and possibly other progenitor cells.
Graphical Abstract
中文翻译：
生理氧张力增强小鼠心脏间充质细胞的能力和功能特性
心脏间充质细胞（CMCs）是一种新发现且有前途的祖细胞，可有效改善心肌梗死后啮齿动物的心脏功能。干细胞/祖细胞通常在大气 O 2压力 (21%) 下培养；然而，心脏中的生理 O 2张力约为 5%，引起了人们对 21% O 2可能因氧化应激而引起毒性的担忧。因此，我们比较了从第 2 代开始以 21% 或 5% O 2培养的小鼠 CMC 。在第 5 代，CMC 经历了严重的缺氧应激（1% O 2持续 24 小时）。与在 21% O 2下培养的 CMC 相比，在 5% O 2下培养在 5 代中持续改善细胞形态，显着减小细胞大小，增加细胞数量，缩短细胞倍增时间，并显着减少缺氧应激后 CMC 释放到培养基中的乳酸脱氢酶。此外，在 5% O 2下培养会增加端粒酶活性和端粒长度，这意味着 21% O 2张力会削弱端粒酶活性，导致端粒缩短和细胞增殖减少。迄今为止，几乎所有针对心脏病细胞疗法的临床前和临床研究都使用大气（21%）O 2来培养细胞。我们的数据挑战了这种范式。我们的结果表明，与 21% O 2、 5% O 2相比张力极大地增强了 CMC 的能力和功能特性。在 5% O 2下增加的增殖率意味着可以以更少的时间和成本实现目标数量的 CMC。此外，由于这种增加的增殖可能在 CMC 移植后在体内继续存在，并且由于在 5% O 2下生长的细胞对严重的缺氧应激有明显的抵抗力，因此在移植到 O 2的心脏疤痕区域后可能能够更好地存活。非常低，在 5% O 2下培养可能会增强 CMC（可能还有其他细胞类型）的修复特性。总之，我们的数据支持用于培养 CMC 和可能的其他祖细胞的方法的变化。
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