Random Mutagenesis, Clonal Events, and Embryonic or Somatic Origin Determine the mtDNA Variant Type and Load in Human Pluripotent Stem Cells.
Random Mutagenesis, Clonal Events, and Embryonic or Somatic Origin Determine the mtDNA Variant Type and Load in Human Pluripotent Stem Cells.
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DOI：
10.1016/j.stemcr.2018.05.007
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发表时间：
2018-07-10
影响因子：
5.9
通讯作者：
Spits C
中科院分区：
文献类型：
--
作者：
Zambelli F;Mertens J;Dziedzicka D;Sterckx J;Markouli C;Keller A;Tropel P;Jung L;Viville S;Van de Velde H;Geens M;Seneca S;Sermon K;Spits C
在这项研究中，我们对人类胚胎和诱导多能干细胞（hESC和hiPSC）及其来源细胞的mtDNA进行了深度测序，发现大多数变体预先存在于用于建立细胞系的细胞中。早期传代的hESC携带很少和低负荷的异质性变体，类似于在卵母细胞和内细胞团中鉴定的那些。随着细胞培养时间的延长，这些变体的数量和异质性负荷增加。对120个早期和晚期传代hESC细胞的研究揭示了单细胞水平mtDNA异质性变体的显着多样性，并且在培养过程中增加的变体总是染色体异常的出现。我们发现，早期传代的hiPSC比hESC携带更高的mtDNA变体负载，单成纤维细胞测序证明其预先存在于源细胞中。最后，我们表明，这些变体在短期分化过程中稳定传播。早期传代的hESC主要携带从胚胎遗传的线粒体DNA变体克隆培养接管导致变体负荷和致病潜力增加hESC和体细胞培养物在线粒体DNA基因型方面是镶嵌的hiPSC由于镶嵌来源的克隆重编程而携带高水平的变体在这项工作中，Spits和同事对hPSC中的线粒体DNA变体进行了深入研究。早期传代的hESC携带很少的低频变体，而在晚期传代的hESC和hiPSC中观察到更高的变体负荷。变异体的存在由品系的体细胞或胚胎起源决定，并通过随机漂移、自发诱变和克隆培养接管来调节。
In this study, we deep-sequenced the mtDNA of human embryonic and induced pluripotent stem cells (hESCs and hiPSCs) and their source cells and found that the majority of variants pre-existed in the cells used to establish the lines. Early-passage hESCs carried few and low-load heteroplasmic variants, similar to those identified in oocytes and inner cell masses. The number and heteroplasmic loads of these variants increased with prolonged cell culture. The study of 120 individual cells of early- and late-passage hESCs revealed a significant diversity in mtDNA heteroplasmic variants at the single-cell level and that the variants that increase during time in culture are always passenger to the appearance of chromosomal abnormalities. We found that early-passage hiPSCs carry much higher loads of mtDNA variants than hESCs, which single-fibroblast sequencing proved pre-existed in the source cells. Finally, we show that these variants are stably transmitted during short-term differentiation. Early-passage hESCs carry mainly mtDNA variants inherited from the embryos Clonal culture takeover results in increased variant load and pathogenic potential hESCs and somatic cell cultures are mosaic in terms of mtDNA genotypes hiPSCs carry high levels of variants due to clonal reprogramming of a mosaic source In this work, Spits and colleagues present an in-depth study of mtDNA variants in hPSCs. Early-passage hESCs carry few low-frequency variants, while higher variant loads are observed in late-passage hESCs and hiPSCs. The presence of variants is determined by the somatic or embryonic origin of the line and modulated by random drift, spontaneous mutagenesis, and clonal culture takeover.
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影响因子：
16.6
作者：
Jacobs, Kurt;Mertzanidou, Afroditi;Spits, Claudia
通讯作者：
Spits, Claudia
影响因子：
5.9
作者：
Avery, Stuart;Hirst, Adam J.;Baker, Duncan;Lim, Chin Yan;Alagaratnam, Sharmini;Skotheim, Rolf I.;Lothe, Ragnhild A.;Pera, Martin F.;Colman, Alan;Robson, Paul;Andrews, Peter W.;Knowles, Barbara B.
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Knowles, Barbara B.
影响因子：
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通讯作者：
Melton, Douglas A.
影响因子：
46.9
作者：
Chambers, Stuart M.;Fasano, Christopher A.;Papapetrou, Eirini P.;Tomishima, Mark;Sadelain, Michel;Studer, Lorenz
通讯作者：
Studer, Lorenz
影响因子：
23.9
作者：
Lamm, Noa;Ben-David, Uri;Kerem, Batsheva
通讯作者：
Kerem, Batsheva