Two Supporting Factors Greatly Improve the Efficiency of Human iPSC Generation

Zhao, Yang; Yin, Xiaolei; Han, Qin; Zhu, Fangfang; Liu, Haisong; Yang, Weifeng; Zhang, Qiang; Xiang, Chengang; Hou, Pingping; Song, Zhihua; Liu, Yanxia; Yong, Jun; Zhang, Pengbo; Cai, Jun; Li, Meng; Li, Honggang; Li, Yanqin; Qu, Xiuxia; Kai, Cui; Zhang, Weiqi; Xiang, T.; Wu, Yetao; Zhao, Yiding; Chun, Liu; Chen, Yu; Yuan, Ke; Lou, Jinning; Ding, Mei; Deng, Hongkui

doi:10.1016/j.stem.2008.10.002

Cited by 429 publications

(366 citation statements)

References 16 publications

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“…Previous reports demonstrated that age‐dependent decline in reprogramming efficiency is correlated with activated intrinsic senescence pathways in cells from old donors (Zhao et al ., 2008; Kawamura et al ., 2009; Li et al ., 2009). Reprogramming SkMs into iPSCs reverses several cellular and molecular characteristics associated with aging, including senescence, telomere truncation, mitochondrial dysfunction, and global changes in gene expression.…”

Section: Discussionmentioning

confidence: 99%

“…After 2–3 weeks, the total number of iPSCs colonies was counted after staining plates for ALP activity following the manufacturer's instructions (Sigma‐Aldrich, St Louis, MO, USA). Based on previously published studies suggesting that exogenous gene silencing might distinguish a pluripotency state, we calculated the number of iPSCs colonies using exogenous mCherry silencing, with morphology resembling mouse ESCs and ALP‐positive staining as the defining criteria (Hotta & Ellis, 2008; Zhao et al ., 2008). Colonies that satisfied these criteria (except ALP staining) were selected and expanded on feeder fibroblasts using the standard procedure.…”

Section: Methodsmentioning

confidence: 99%

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Blockade of senescence‐associated micro RNA ‐195 in aged skeletal muscle cells facilitates reprogramming to produce induced pluripotent stem cells.

et al. 2015

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SummaryThe low reprogramming efficiency in cells from elderly patients is a challenge that must be overcome. Recently, it has been reported that senescence‐associated microRNA (miR)‐195 targets Sirtuin 1 (SIRT1) to advance cellular senescence. Thus, we hypothesized that a blockade of miR‐195 expression could improve reprogramming efficiency in old skeletal myoblasts (SkMs). We found that miR‐195 expression was significantly higher in old SkMs (24 months) isolated from C57BL/6 mice as compared to young SkMs (2 months, 2.3‐fold). Expression of SIRT1 and telomerase reverse transcriptase (TERT) was downregulated in old SkMs, and transduction of old SkMs with lentiviral miR‐195 inhibitor significantly restored their expression. Furthermore, quantitative in situ hybridization analysis demonstrated significant telomere elongation in old SkMs transduced with anti‐miR‐195 (1.7‐fold increase). It is important to note that blocking miR‐195 expression markedly increased the reprogramming efficiency of old SkMs as compared to scramble (2.2‐fold increase). Transduction of anti‐miR‐195 did not alter karyotype or pluripotency marker expression. Induced pluripotent stem cells (iPSCs) from old SkMs transduced with anti‐miR‐195 successfully formed embryoid bodies that spontaneously differentiated into three germ layers, indicating that deletion of miR‐195 does not affect pluripotency in transformed SkMs. In conclusion, this study provided novel evidence that the blockade of age‐induced miR‐195 is a promising approach for efficient iPSC generation from aging donor subjects, which has the potential for autologous transplantation of iPSCs in elderly patients.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Blockade of senescence‐associated micro RNA ‐195 in aged skeletal muscle cells facilitates reprogramming to produce induced pluripotent stem cells.

et al. 2015

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“…Only a few cells can overcome this barrier and become iPSCs. A number of studies demonstrated that when key components (such as p53 and p21) of the DNA damage pathway were deleted, the percentage of iPS cell generation significantly increased (Zhao et al, 2008a;Banito et al, 2009;Hong et al, 2009;Kawamura et al, 2009;Li et al, 2009;Marión et al, 2009;Utikal et al, 2009b). Suppression of the p53 pathway may compromise a cell's genome stability.…”

Section: Overcoming the Dna Damage Responsementioning

confidence: 99%

“…As the DNA damage pathway poses a major barrier towards regaining pluripotency (Zhao et al, 2008b;Banito et al, 2009;Hong et al, 2009;Kawamura et al, 2009;Li et al, 2009;Marión et al, 2009;Utikal et al, 2009b), chemical compounds that alleviate the stress of DNA damage response may help cells to overcome this barrier. To this end, Vitamin C has been shown to significantly increase the formation of both mouse and human iPSC by suppressing p53 induced cell senescence (Esteban et al, 2010).…”

Section: Chemical Compoundsmentioning

confidence: 99%

Mechanism and methods to induce pluripotency

Wang

2011

Protein Cell

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Pluripotent stem cells are able to self-renew indefinitely and differentiate into all types of cells in the body. They can thus be an inexhaustible source for future cell transplantation therapy to treat degenerative diseases which currently have no cure. However, non-autologous cells will cause immune rejection. Induced pluripotent stem cell (iPSC) technology can convert somatic cells to the pluripotent state, and therefore offers a solution to this problem. Since the first generation of iPSCs, there has been an explosion of relevant research, from which we have learned much about the genetic networks and epigenetic landscape of pluripotency, as well as how to manipulate genes, epigenetics, and microRNAs to obtain iPSCs. In this review, we focus on the mechanism of cellular reprogramming and current methods to induce pluripotency. We also highlight new problems emerging from iPSCs. Better understanding of the fundamental mechanisms underlying pluripotenty and refining the methodology of iPSC generation will have a significant impact on future development of regenerative medicine.

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“…Zhao et al, 2008;Hong et al, 2009;Kawamura et al, 2009;Li et al, 2009a;Marion et al, 2009;Utikal et al, 2009 Signaling pathway TGF-β Inhibition of TGF-β signaling promotes the completion of reprogramming through induction of the transcription factor Nanog. Any one of TGF-β inhibitor E-616452 ("RepSox") or E-616451 can replace Sox2 Ichida et al, 2009 Reprogramming is the iterative replacement of the original transcriptional program present within the differentiated starting cell with the program present within a pluripotential cell: a key step is the silencing of the molecular program of the donor cell.…”

Section: Enhancing Reprogramming Efficiency By Alternative Factorsmentioning

confidence: 99%

Overcoming barriers to the clinical utilization of iPSCs: reprogramming efficiency, safety and quality

et al. 2012

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Differentiated cells can be reprogrammed into pluripotent stem cells, known as "induced pluripotent stem cells" (iPSCs), through the overexpression of defined transcription factors. The creation of iPSC lines has opened new avenues for patient-specific cell replacement therapies for regenerative medicine. However, the clinical utilization of iPSCs is largely impeded by two limitations. The first limitation is the low efficiency of iPSCs generation from differentiated cells. The second limitation is that many iPSC lines are not authentically pluripotent, as many cell lines inefficiently differentiate into differentiated cell types when they are tested for their ability to complement embryonic development. Thus, the "quality" of iPSCs must be increased if they are to be differentiated into specialized cell types for cell replacement therapies. Overcoming these two limitations is paramount to facilitate the widespread employment of iPSCs for therapeutic purposes. Here, we summarize recent progress made in strategies enabling the efficient production of high-quality iPSCs, including choice of reprogramming factors, choice of target cell type, and strategies to improve iPSC quality.

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Two Supporting Factors Greatly Improve the Efficiency of Human iPSC Generation

Cited by 429 publications

References 16 publications

Blockade of senescence‐associated micro RNA ‐195 in aged skeletal muscle cells facilitates reprogramming to produce induced pluripotent stem cells.

Blockade of senescence‐associated micro RNA ‐195 in aged skeletal muscle cells facilitates reprogramming to produce induced pluripotent stem cells.

Mechanism and methods to induce pluripotency

Overcoming barriers to the clinical utilization of iPSCs: reprogramming efficiency, safety and quality

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