Efficient generation of integration-free ips cells from human adult peripheral blood using BCL-XL together with Yamanaka factors.
Efficient generation of integration-free ips cells from human adult peripheral blood using BCL-XL together with Yamanaka factors.
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DOI：
10.1371/journal.pone.0064496
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Zhang XB
中科院分区：
文献类型：
--
作者：
Su RJ;Baylink DJ;Neises A;Kiroyan JB;Meng X;Payne KJ;Tschudy-Seney B;Duan Y;Appleby N;Kearns-Jonker M;Gridley DS;Wang J;Lau KH;Zhang XB
从最容易获得的来源-外周血中高效地产生无整合诱导多能干细胞(IPSC)的能力有可能加速基于IPSC的治疗的进展。我们已经成功地从脐带血(CB)CD34+细胞和改良的基于ORIP/EBNA1的外体载体(EV)中，利用强大的脾病灶形成病毒(SFFV)长末端重复序列(LTR)启动子，从脐血CD34+细胞中获得了无整合的IPSCs。在这里，我们表明山中因子(OCT4、SOX2、MYC和KLF4)表达的EV也可以将成人外周血单个核细胞(PBMNC)重新编程为多能细胞，但效率非常低。我们发现，包含BCL-XL可将重新编程效率提高约10倍。此外，CD3−/CD19IPSC或去T/B细胞的单核细胞培养4-6天后，每毫升外周血可产生20-30个−集落，这一效率显著高于先前报道的结果。外周血间充质干细胞表达多能标志物，可形成畸胎瘤，在体外可被诱导分化为间充质干细胞、心肌细胞和肝细胞。结合使用，我们的优化因子组合和重新编程策略可以从成人PB高效地生成无整合的IPSCs。这一发现在IPSC银行、疾病建模和再生医学中具有潜在的应用价值。
The ability to efficiently generate integration-free induced pluripotent stem cells (iPSCs) from the most readily available source—peripheral blood—has the potential to expedite the advances of iPSC-based therapies. We have successfully generated integration-free iPSCs from cord blood (CB) CD34+ cells with improved oriP/EBNA1-based episomal vectors (EV) using a strong spleen focus forming virus (SFFV) long terminal repeat (LTR) promoter. Here we show that Yamanaka factors (OCT4, SOX2, MYC, and KLF4)-expressing EV can also reprogram adult peripheral blood mononuclear cells (PBMNCs) into pluripotency, yet at a very low efficiency. We found that inclusion of BCL-XL increases the reprogramming efficiency by approximately 10-fold. Furthermore, culture of CD3−/CD19− cells or T/B cell-depleted MNCs for 4–6 days led to the generation of 20–30 iPSC colonies from 1 ml PB, an efficiency that is substantially higher than previously reported. PB iPSCs express pluripotency markers, form teratomas, and can be induced to differentiate in vitro into mesenchymal stem cells, cardiomyocytes, and hepatocytes. Used together, our optimized factor combination and reprogramming strategy lead to efficient generation of integration-free iPSCs from adult PB. This discovery has potential applications in iPSC banking, disease modeling and regenerative medicine.
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影响因子：
1.2
作者：
Bai, Hao;Chen, Kang;Gao, Yong-Xing;Arzigian, Melanie;Xie, Yin-Liang;Malcosky, Christopher;Yang, Yong-Guang;Wu, Wen-Shu;Wang, Zack Z.
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影响因子：
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作者：
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通讯作者：
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影响因子：
20.3
作者：
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通讯作者：
Naldini, L
影响因子：
23.9
作者：
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通讯作者：
Jaenisch, Rudolf