Generation of induced pluripotent stem cells from human cord blood cells with only two factors: Oct4 and Sox2

Giorgetti, Alessandra; Montserrat, Núria; Rodríguez-Pizá, Ignacio; Azqueta, Carmen; Veiga, Anna; Belmonte, Juan Carlos Izpisúa

doi:10.1038/nprot.2010.16

Cited by 94 publications

(65 citation statements)

References 13 publications

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“…Instead, we found that coculture with irradiated human foreskin fibroblasts (i.e., HFF-1 cells) in the presence of bFGF (23) (Fig. 1A) were permissive for the survival of Sox2-transduced CB cells.…”

Section: Resultsmentioning

confidence: 56%

Cord blood-derived neuronal cells by ectopic expression of Sox2 and c-Myc

Giorgetti

Marchetto

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The finding that certain somatic cells can be directly converted into cells of other lineages by the delivery of specific sets of transcription factors paves the way to novel therapeutic applications. Here we show that human cord blood (CB) CD133 + cells lose their hematopoietic signature and are converted into CB-induced neuronal-like cells (CB-iNCs) by the ectopic expression of the transcription factor Sox2, a process that is further augmented by the combination of Sox2 and c-Myc. Gene-expression analysis, immunophenotyping, and electrophysiological analysis show that CBiNCs acquire a distinct neuronal phenotype characterized by the expression of multiple neuronal markers. CB-iNCs show the ability to fire action potentials after in vitro maturation as well as after in vivo transplantation into the mouse hippocampus. This system highlights the potential of CB cells and offers an alternative means to the study of cellular plasticity, possibly in the context of drug screening research and of future cell-replacement therapies.neurons | reprogramming | stem cells T he fate of adult somatic cells is not fixed rigidly and can be reprogrammed by experimental manipulation. The generation of induced pluripotent stem cells (iPSCs) represents the most dramatic evidence that the epigenome of somatic cells are remarkably plastic (1). Recently, it has been reported that fibroblasts can be converted by ectopic expression of defined transcription factors into postmitotic neurons (2-9), neural progenitors (10, 11), and self-renewing neural stem cells (NSCs) (12, 13). However, most reported methods rely on the use of multiple transcription factors and the use of fibroblasts as donor cells.Here we investigate if cord blood (CB) stem cells can be induced to acquire a neuronal phenotype by using only one transcription factor. It has been previously shown that stem cell populations are more amenable to reprogramming than other somatic cells, probably as a result of their preexisting epigenetic state (14,15). Moreover, because of their biological characteristics and availability, CB cells as a donor cell type could offer clear logistic advantages vs. other adult somatic cell types (16,17). These cells can be collected without any risk for the donor and are young cells carrying minimal somatic mutations.Strikingly, we show that forced expression of Sox2 is sufficient to convert CB CD133 + cells into induced neuronal progenitor (NP)-like cells, a conversion process that is augmented by the coexpression of c-Myc. Sox2 is highly expressed in adult NSCs, is one of the earliest functional markers of neuroectodermal specification in the embryo, and plays a key role in the neural lineage specification (18)(19)(20). We show that Sox2-transduced CB cells acquire a distinct neuronal morphology and express multiple neuronal markers in vitro, and that they can be expanded for as many as 25 passages when cultured in permissive condition culture. Moreover, we show that CB-induced neuronal-like cells (CB-iNCs) are able to differentiate in vitro and ...

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Section: Resultsmentioning

confidence: 56%

Cord blood-derived neuronal cells by ectopic expression of Sox2 and c-Myc

Giorgetti

Marchetto

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…þ cord blood cells (expressing pluripotency markers and capable of forming EB and teratomas) were generated in two weeks whereas no colonies were obtained with control keratinocytes or fibroblasts using the two factors only (Giorgetti et al 2009(Giorgetti et al , 2010. CD133 þ cord blood cells and foetal NSCs already have a baseline level expression of Oct-4 and/or Sox2 and that, along with a generally more permissive chromatin organization, might be the key to their greater reprogramming efficiency compared with adult cells.…”

Section: Use Of Fscs For Reprogrammingmentioning

confidence: 99%

Biological characteristics of stem cells from foetal, cord blood and extraembryonic tissues

Abdulrazzak

Moschidou

Jones

et al. 2010

J. R. Soc. Interface.

130

118

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Foetal stem cells (FSCs) can be isolated during gestation from many different tissues such as blood, liver and bone marrow as well as from a variety of extraembryonic tissues such as amniotic fluid and placenta. Strong evidence suggests that these cells differ on many biological aspects such as growth kinetics, morphology, immunophenotype, differentiation potential and engraftment capacity in vivo. Despite these differences, FSCs appear to be more primitive and have greater multi-potentiality than their adult counterparts. For example, foetal blood haemopoietic stem cells proliferate more rapidly than those found in cord blood or adult bone marrow. These features have led to FSCs being investigated for pre-and post-natal cell therapy and regenerative medicine applications. The cells have been used in pre-clinical studies to treat a wide range of diseases such as skeletal dysplasia, diaphragmatic hernia and respiratory failure, white matter damage, renal pathologies as well as cancers. Their intermediate state between adult and embryonic stem cells also makes them an ideal candidate for reprogramming to the pluripotent status.

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“…Figure 5 Relative gene expression levels of Oct4, Sox2, and Nanog in human fibroblasts, keratinocytes, CD133 þ cells from cord blood, and NSCs. The relative gene expression levels in fibroblasts, keratinocytes, and CD133 þ cells compared with hESCs were calculated from the data reported by Giorgetti et al 85 and Page et al 86 The relative gene expression levels in human NSCs compared with hESCs were used from the data reported by Kim et al 13 NSCs endogenously express Sox2, Klf4, and/or c-Myc to some extent, as was discussed in the previous section ( Figure 5). Therefore, Kim et al reprogrammed human fetal NSCs into hiPSCs by the transduction of only Oct4.…”

Section: Small Molecules Can Induce Human Ipscs In Combination With Amentioning

confidence: 99%

“…Some examples of the gene expression levels of Oct4, Sox2, and Nanog in human fibroblasts, keratinocytes, CD133 þ cells from cord blood, and NSCs are described in Figure 5. 13,85,86 Several researchers tried to use endogenously expressing Sox2, Klf4, and/or c-Myc and to generate iPSCs with fewer numbers of pluripotent TFs. Kim et al succeeded in preparing miPSCs from mouse NSCs using two exogenous TFs (Oct4 and Klf4 or Oct4 and c-Myc).…”

Section: Small Molecules Can Replace Several Tfs During Mouse Ipsc Rementioning

confidence: 99%

Generation of pluripotent stem cells without the use of genetic material

Higuchi

Ling

Kumar

et al. 2015

Laboratory Investigation

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Induced pluripotent stem cells (iPSCs) provide a platform to obtain patient-specific cells for use as a cell source in regenerative medicine. Although iPSCs do not have the ethical concerns of embryonic stem cells, iPSCs have not been widely used in clinical applications, as they are generated by gene transduction. Recently, iPSCs have been generated without the use of genetic material. For example, protein-induced PSCs and chemically induced PSCs have been generated by the use of small and large (protein) molecules. Several epigenetic characteristics are important for cell differentiation; therefore, several small-molecule inhibitors of epigenetic-modifying enzymes, such as DNA methyltransferases, histone deacetylases, histone methyltransferases, and histone demethylases, are potential candidates for the reprogramming of somatic cells into iPSCs. In this review, we discuss what types of small chemical or large (protein) molecules could be used to replace the viral transduction of genes and/or genetic reprogramming to obtain human iPSCs.

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Generation of induced pluripotent stem cells from human cord blood cells with only two factors: Oct4 and Sox2

Cited by 94 publications

References 13 publications

Cord blood-derived neuronal cells by ectopic expression of Sox2 and c-Myc

Cord blood-derived neuronal cells by ectopic expression of Sox2 and c-Myc

Biological characteristics of stem cells from foetal, cord blood and extraembryonic tissues

Generation of pluripotent stem cells without the use of genetic material

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