Generation of Induced Pluripotent Stem Cells by Efficient Reprogramming of Adult Bone Marrow Cells

Kunisato, Atsushi; Wakatsuki, Mariko; Kodama, Yuuki; Shinba, Haruna; Ishida, Isao; Nagao, Kazuharu

doi:10.1089/scd.2009.0149

Cited by 64 publications

(36 citation statements)

References 46 publications

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“…+ cord blood cells, bone marrow mononuclear cells (BM-MNCs), and adipose tissuederived mesenchymal stem cells (AT-MSCs), have been tested and evaluated for cell reprogramming (Takahashi and Yamanaka, 2006;Kim et al, 2009;Sun et al, 2009;Aasen and Belmonte, 2010;Kunisato et al, 2010;Takenaka et al, 2010). Compared with other cell types, AT-MSCs have been shown to be compatible with embryonic stem (ES) cell culture medium and used as feeder layers for ES cell growth.…”

Section: Introductionmentioning

confidence: 99%

Reprogramming Adipose Tissue-Derived Mesenchymal Stem Cells into Pluripotent Stem Cells by a Mutant Adeno-Associated Viral Vector

Chen

Hamazaki

et al. 2014

Human Gene Therapy Methods

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Induced pluripotent stem (iPS) cells have great potential for personalized regenerative medicine. Although several different methods for generating iPS cells have been reported, improvement of safety and efficiency is imperative. In this study, we tested the feasibility of using a triple tyrosine mutant AAV2 (Y444 + 500 + 730F) vector, designated AAV2.3m, to generate iPS cells. We developed a polycistronic rAAV2.3m vector expressing three reprogramming factors, Klf4, Oct4, and Sox2, and then used this vector to infect mouse adipose-derived mesenchymal stem cells (AT-MSCs) to induce the generation of iPS cells. We demonstrated that (1) the triple tyrosine mutant AAV2 vector is able to reprogram mouse adult adipose tissue-derived stem cells into the pluripotent state. Those rAAV2.3m-derived iPS (rAAV2.3m-iPS) cells express endogenous pluripotencyassociated genes including Oct4, Sox2, and SSEA-1, and form teratomas containing multiple tissues in vivo; (2) c-myc, an oncogene, is dispensable in rAAV2.3m-mediated cellular reprogramming; and (3) transgene expression is undetectable after reprogramming, whereas vector DNA is detectable, indicating that transgenes are silenced. These results indicated the rAAV vector may have some advantages in generating iPS cells.

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Section: Introductionmentioning

confidence: 99%

Reprogramming Adipose Tissue-Derived Mesenchymal Stem Cells into Pluripotent Stem Cells by a Mutant Adeno-Associated Viral Vector

Chen

Hamazaki

et al. 2014

Human Gene Therapy Methods

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“…The comparison of different cell types derived from the same inbred Various other starting somatic cell populations have been used for iPS induction in the mouse, including transgenic mouse strain also negates any possible genotype effects present when trying to draw conclusions adult stomach and liver cells (2), embryonic or newborn neural progenitor cells (9,14), pancreatic β-cells (29), from different studies. While drug selection has been used extensively to generate iPS colonies, timing of drug mature B lymphocytes (10), and bone marrow hematopoietic cells (15). An important observation from these selection has been reported to greatly influence iPS colony numbers and the level of reprogramming attained studies is that the somatic cell type chosen had a significant influence on the efficiency of iPS generation and (4, 18,19).…”

Section: Introductionmentioning

confidence: 99%

The Efficient Generation of Induced Pluripotent Stem (iPS) Cells from Adult Mouse Adipose Tissue-Derived and Neural Stem Cells

et al. 2010

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Ectopic expression of key reprogramming transgenes in somatic cells enables them to adopt the characteristics of pluripotency. Such cells have been termed induced pluripotent stem (iPS) cells and have revolutionized the field of somatic cell reprogramming, as the need for embryonic material is obviated. One of the issues facing both the clinical translation of iPS cell technology and the efficient derivation of iPS cell lines in the research laboratory is choosing the most appropriate somatic cell type for induction. In this study, we demonstrate the direct reprogramming of a defined population of neural stem cells (NSCs) derived from the subventricular zone (SVZ) and adipose tissue-derived cells (ADCs) from adult mice using retroviral transduction of the Yamanaka factors Oct4, Sox2, Klf4, and c-Myc, and compared the results obtained with a mouse embryonic fibroblast (mEF) control. We isolated mEFs, NSCs, and ADCs from transgenic mice, which possess a GFP transgene under control of the Oct4 promoter, and validated GFP expression as an indicator of reprogramming. While transduction efficiencies were not significantly different among the different cell types (mEFs 68.70 ± 2.62%, ADCs 70.61 ± 15.4%, NSCs, 68.72 ± 3%, p = 0.97), the number of GFP-positive colonies and hence the number of reprogramming events was significantly higher for both NSCs (13.50 ± 4.10 colonies, 0.13 ± 0.06%) and ADCs (118.20 ± 38.28 colonies, 1.14 ± 0.77%) when compared with the mEF control (3.17 ± 0.29 colonies, 0.03 ± 0.005%). ADCs were most amenable to reprogramming with an 8-and 38-fold greater reprogramming efficiency than NSCs and mEFs, respectively. Both NSC iPS and ADC iPS cells were demonstrated to express markers of pluripotency and could differentiate to the three germ layers, both in vitro and in vivo, to cells representative of the three germ lineages. Our findings confirm that ADCs are an ideal candidate as a readily accessible somatic cell type for high efficiency establishment of iPS cell lines.Key words: Induced pluripotent stem (iPS) cells; Reprogramming; Adipose-derived cells; Neural stem cells INTRODUCTIONproach, ethical and logistical concerns have largely diminished their utility [reviewed in Hochedlinger and Jaenisch (11)]. Pluripotency describes the ability of a cell to differentiate into cells of the three germ lineages. This property In 2006, the pioneering work of Takahashi and Yamanaka showed that it was possible to directly reprois usually restricted to a few cell types, including cells of the inner cell mass of a preimplanted blastocyst and gram mouse adult fibroblasts into pluripotent-like cells, without the use of embryonic material (36). The cells their in vitro counterparts, ES cells. While somatic cells have a restricted ability to differentiate during normal were termed induced pluripotent stem (iPS) cells and were generated by the retroviral transduction of four development, under certain experimental conditions they can be induced to revert to a pluripotent state. This protranscription factors: Oct4, So...

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“…In this case, our choice of using identical starting parental cells, while having the benefit of creating a system with genetically identical cells, in addition, introduced a potential bias into our comparison. This possibility was further examined by examining the gene expression profile of BM iPSCs [38]. Indeed, there was a decrease in expression of differentiationassociated genes in BM iPSCs that are associated with the parental bone marrow cells that are not shared with fibroblasts.…”

Section: Discussionmentioning

confidence: 99%

“…We also found that both cell types commonly upregulated the glycolysis pathway, although through different genes in each case. We also examined 2 other published microarray data sets generated from iPSCs that were derived either by using different cocktails of reprogramming factors [37] or different starting cells [38] to determine the global validity of our comparisons and found very similar results, suggesting that the similarity of iPSCs and OF is independent of both Myc and fibroblastic parental cells. There were some key differences, however, between iPSCs and OF.…”

Section: Introductionmentioning

confidence: 94%

“…To ascertain the potential influence of parental cell type on loss of differentiation-associated gene expression during the iPSC formation, we also examined a data set published on iPSCs derived using bone marrow mononuclear cells (BM iPSCs) from adult mice as starting material [38]. We generated lists of genes at least 1.5-fold downregulated in BM iPSCs as compared to their starting cells, and then compared that list of genes to those genes downregulated in our OF and mESCs compared to parental MEFs (Supplementary Fig.…”

Section: Riggs Et Almentioning

confidence: 99%

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Induced Pluripotency and Oncogenic Transformation Are Related Processes

Riggs

Barrilleaux

Varlakhanova

et al. 2013

Stem Cells and Development

105

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Induced pluripotent stem cells (iPSCs) have the potential for creating patient-specific regenerative medicine therapies, but the links between pluripotency and tumorigenicity raise important safety concerns. More specifically, the methods employed for the production of iPSCs and oncogenic foci (OF), a form of in vitro produced tumor cells, are surprisingly similar, raising potential concerns about iPSCs. To test the hypotheses that iPSCs and OF are related cell types and, more broadly, that the induction of pluripotency and tumorigenicity are related processes, we produced iPSCs and OF in parallel from common parental fibroblasts. When we compared the transcriptomes of these iPSCs and OF to their parental fibroblasts, similar transcriptional changes were observed in both iPSCs and OF. A significant number of genes repressed during the iPSC formation were also repressed in OF, including a large cohort of differentiation-associated genes. iPSCs and OF shared a limited number of genes that were upregulated relative to parental fibroblasts, but gene ontology analysis pointed toward monosaccharide metabolism as upregulated in both iPSCs and OF. iPSCs and OF were distinct in that only iPSCs activated a host of pluripotency-related genes, while OF activated cellular damage and specific metabolic pathways. We reprogrammed oncogenic foci (ROF) to produce iPSC-like cells, a process dependent on Nanog. However, the ROF had reduced differentiation potential compared to iPSC, suggesting that oncogenic transformation leads to cellular changes that impair complete reprogramming. Taken together, these findings support a model in which OF and iPSCs are related, yet distinct cell types, and in which induced pluripotency and induced tumorigenesis are similar processes.

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Generation of Induced Pluripotent Stem Cells by Efficient Reprogramming of Adult Bone Marrow Cells

Cited by 64 publications

References 46 publications

Reprogramming Adipose Tissue-Derived Mesenchymal Stem Cells into Pluripotent Stem Cells by a Mutant Adeno-Associated Viral Vector

Reprogramming Adipose Tissue-Derived Mesenchymal Stem Cells into Pluripotent Stem Cells by a Mutant Adeno-Associated Viral Vector

The Efficient Generation of Induced Pluripotent Stem (iPS) Cells from Adult Mouse Adipose Tissue-Derived and Neural Stem Cells

Induced Pluripotency and Oncogenic Transformation Are Related Processes

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