The mutational impact of culturing human pluripotent and adult stem cells
The mutational impact of culturing human pluripotent and adult stem cells
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DOI：
10.1038/s41467-020-16323-4
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发表时间：
2020-05-19
影响因子：
16.6
通讯作者：
Cuppen, Edwin
中科院分区：
文献类型：
--
作者：
Kuijk, Ewart;Jager, Myrthe;Cuppen, Edwin
在体外培养过程中获得的遗传变化对干细胞在再生医学中的成功应用构成了风险。为了评估培养诱导的遗传风险，我们通过全基因组测序确定了个体人类干细胞的所有突变。个体多能干细胞、肠干细胞和肝干细胞每倍增一次分别累积3.50.5、7.2 +/- 1.1和8.3 +/- 3.6个碱基替换。成体干细胞体外的年突变积累速率比体内的突变积累速率高近40倍。突变特征分析表明，体外诱导突变是由氧化应激引起的。降低培养中的氧张力可降低突变负荷。我们使用突变率、光谱和基因组分布来模拟人类干细胞在典型的体外扩增、操作或筛选实验中致癌突变的积累。我们的研究提供了经验定义的参数来评估干细胞治疗的突变风险。
Genetic changes acquired during in vitro culture pose a risk for the successful application of stem cells in regenerative medicine. To assess the genetic risks induced by culturing, we determined all mutations in individual human stem cells by whole genome sequencing. Individual pluripotent, intestinal, and liver stem cells accumulate 3.50.5, 7.2 +/- 1.1 and 8.3 +/- 3.6 base substitutions per population doubling, respectively. The annual in vitro mutation accumulation rate of adult stem cells is nearly 40-fold higher than the in vivo mutation accumulation rate. Mutational signature analysis reveals that in vitro induced mutations are caused by oxidative stress. Reducing oxygen tension in culture lowers the mutational load. We use the mutation rates, spectra, and genomic distribution to model the accumulation of oncogenic mutations during typical in vitro expansion, manipulation or screening experiments using human stem cells. Our study provides empirically defined parameters to assess the mutational risk of stem cell based therapies.