Retracted

Stimulus-triggered fate conversion of somatic cells into pluripotency

Article (PDF Available)inNature 505(7485):641-7 · January 2014with 8,798 Reads
DOI: 10.1038/nature12968 · Source: PubMed
Abstract
Here we report a unique cellular reprogramming phenomenon, called stimulus-triggered acquisition of pluripotency (STAP), which requires neither nuclear transfer nor the introduction of transcription factors. In STAP, strong external stimuli such as a transient low-pH stressor reprogrammed mammalian somatic cells, resulting in the generation of pluripotent cells. Through real-time imaging of STAP cells derived from purified lymphocytes, as well as gene rearrangement analysis, we found that committed somatic cells give rise to STAP cells by reprogramming rather than selection. STAP cells showed a substantial decrease in DNA methylation in the regulatory regions of pluripotency marker genes. Blastocyst injection showed that STAP cells efficiently contribute to chimaeric embryos and to offspring via germline transmission. We also demonstrate the derivation of robustly expandable pluripotent cell lines from STAP cells. Thus, our findings indicate that epigenetic fate determination of mammalian cells can be markedly converted in a context-dependent manner by strong environmental cues.

Supplementary resources

  • ... The national scandal in Japan involving STAP cells developed around an original research article and its companion letter published in Nature, one of the most prom- inent journals in science, on 30 January 2014. Obokata et al. (2014aObokata et al. ( , 2014b) reported development of a new method to reprogram differentiated somatic cells of a mouse and induce them to be pluripotent, that is, capable of becoming any type of cell in the body. Two days before publication, the first author of the publications, Obokata, and her colleagues at the CDB announced at a press conference that they had successfully created what they named stimulus-triggered acquisition of pluripotency (STAP) cells and had established stable cell lines from them, which they called STAP stem cells. ...
    ... The pioneers of these methods, John B. Gurdon and Shinya Yamanaka, respectively, jointly received the Nobel Prize in Physiology or Medicine in 2012. Yet, as described in the research article ( Obokata et al. 2014b), the new method differed in that STAP was understood to be a phenomenon that occurs naturally in organisms, while cloning and genetic manipulation are not. This biological phenomenon is known to happen in plants, and the authors claimed that they discovered it happens equally in animals. ...
    ... This discourse was not a mere consequence of media (mis)representation. While STAP cells were compared mainly with ES cells in the original research article ( Obokata et al. 2014b), their superiority to iPS cells was emphasized in the press conference (Suda 2014). First, it was highlighted that the reprogramming of somatic cells could be done more effectively and faster with the new method than with the "old" reprogramming technique. ...
    Article
    On 28 January 2014, Haruko Obokata and her colleagues held a press conference regarding their new method of producing stem cells. The cells, named STAP (stimulus-triggered acquisition of pluripotency) cells, were of considerable interest not only for stem cell scientists but also for the wider society in Japan because both its government and citizens enjoyed the international reputation earned for the country by Shinya Yamanaka’s earlier success in developing a novel technique of cell reprogramming. However, it was soon pointed out that the data in the research article seemed fabricated and was hence suggested that their claims lacked scientific credibility. What was initially considered another triumph of Japanese stem cell research thus resulted in a major national scandal. Instead of seeing this casemerely as one of scientificmisconduct, this article examines it as a windowinto the local culture of stemcell research and argues that the socio-institutional background ofObokata’swork incited the researchers involved to infer doability of STAP cell research. The problem with this inference surfaced after those who did not share the culture challenged the robustness of her work, suggesting some cultures of science may be more vulnerable to scientific misconduct than others.
  • ... In January 2014 Haruko Obokata published two breakthrough articles in "Nature" in which she introduced a simple method of STAP ("stimulus-triggered acquisition of pluripotency") [14,15]. Obokata revealed that "strong external stimuli such as a transient low-pH stressor reprogrammed mammalian somatic cells, resulting in the generation of pluripotent cells" [15]. ...
    ... In January 2014 Haruko Obokata published two breakthrough articles in "Nature" in which she introduced a simple method of STAP ("stimulus-triggered acquisition of pluripotency") [14,15]. Obokata revealed that "strong external stimuli such as a transient low-pH stressor reprogrammed mammalian somatic cells, resulting in the generation of pluripotent cells" [15]. However, soon after publication disturbing allegations emerged. ...
    Conference Paper
    Full-text available
    Cell cultures are believed to be a powerful research tool, finding its applications in various fields of science. Ever since Alexis Carrel, the Nobel Prize laureate, established cell culture of chick heart fibroblasts [1], the scientific race for the far-reaching discoveries has begun. The problem is, that Carrel had proven “beyond doubt” that his cells can live for almost infinite period of time, which was impossible to replicate in other laboratories. The addition of fresh cells appeared to be the answer to the mystery [2,3], but whether it had been a simple mistake or deliberate fraud remains an open question. Although Carrel’s misleading conclusion was subsequently exposed by other scientists, the example of the world-famous deception by Haruko Obokata and her Stap cells or the case of Hwang Woo Suk who has supposedly cloned human embryonic stem cells show that a large threat can be related to unreliable and careless wet lab research [4]. This work is an overview of the history and practices in the cell culture laboratories. We will analyse the successes and failures in this scientific field and based on that and our own experience, we will present the pros and cons of using this model in modern research laboratories. ACKNOWLEDGEMENTS: This work was supported by funds from the project STM.A040.17.039. REFERENCES: [1] A. Carrel: On the permanent life of tissues outside of the organism. J. Exp. Med. vol. 15, no. 5, pp. 516–28, 1912. [2] J.A. Witkowski: Dr. Carrel’s immortal cells. Med. Hist. vol. 24, no. 2, pp. 129-42, 1980. [3] J. Witkowski: The myth of cell immortality. Trends Biochem. Sci. vol. 10, no. 7, pp. 258-260, 1985. [4] J. Rasko, C. Power: What pushes scientists to lie? The disturbing but familiar story of Haruko Obokata, Guardian, Wed 18 Feb 2015.
  • ... If MSs had recorded their experiments in these situations, many problems would have been alleviated. Moreover, if electronic recording had been performed as a rule among the scientists and experiments related to the stimulus-triggered acquisition of pluripotency cells, the subsequent STAP Cells Scandal may have been prevented [7][8][9] . As mentioned above, recording systems are costly ; therefore, we propose that the Ministry of Health, Labor & Welfare lead the initiative to distribute recording systems among MSs, as well as subsidize universities and colleges to support the effort. ...
    Article
    Medical specialists (scientists, doctors, nurses, pharmacists, etc.) are under strict scrutiny in contemporary society as they are held to a high standard and their crimes are loudly broadcasted through the mass media. In this study, we introduce three cases including the Novartis Case, involving a pharmaceutical company employee, the Mazindol Case involving a doctor, and the Muscle Relaxant Case involving a practical nurse. We analyzed each case, studying the risk and protective factors, to determine what caused the above-mentioned medical specialists to commit medical malpractice. In addition to identification and discussion of the key risk factors involved in each case, we propose prevention measures for medical specialists, particularly scientists, in order to avoid similar acts of misconduct in the future. We conclude that additional education and transparency regarding work performance are necessary to prevent such violations. To eliminate doubt, we recommend that electronic recording devices be used by medical specialists during experimentation.
  • ... Сучасні експериментальні дослідження на субклітинному рівні показали, що вплив стресорів, зумовлюючи пошкодження тканин різної локалізації, здатний трансформовувати зрілі соматичні клітини (паранефральний епітелій, клітини проксимальних звивистих канальців нефрона, епітелій трахеї, шлунка) в стовбурові плюрипотентні, ембріональні, цей процес отримав назву "дедиференціація" [Obokata H. et al., 2014]. Такі спостереження розширюють уявлення про регенераторний потенціал організму та відкривають нові горизонти для лікування та профілактики багатьох хвороб. ...
  • Article
    Important and potentially useful findings in the sciences are under more intense public scrutiny now more than ever. Other researchers in the field dive into replicating and expanding the findings while the media swamps the community and the public with peripheral reporting and analyses. How should authors and the hosting/funding institutions respond when other workers in the field could not reproduce or replicate their published results? To illustrate the importance of author-initiated and institution-driven investigations in response to outcries of research irreproducibility, I draw on comparisons between three recent and well-publicized cases in the life sciences: betatrophin, STAP cells, and NgAgo. Swift, transparent responses and investigations facilitate activation of the self-correcting mechanism of science and are likely also critical in preserving the community’s resources, public trust, and the reputation of the institutions and individuals concerned. Operational guidelines for “author and institutional responses” towards external reports of irreproducibility should therefore be in place for all research intensive institutions.
  • Chapter
    In this chapter, the focus is turned from the hypothetical and anticipatory perspective of stem cell-based clinical translation to the “realities” of laboratory life with human pluripotent stem cells. Here practical challenges of instrumentalizing the artificially created iPS cell lines and their volatile and fickle vitality become poignant. The chapter describes the discovery of the revolutionary method of cellular reprogramming, followed by the analyses on what is required of biomedical craftwork to make use of the new biological tool and how this work is conditioned by and dependent on the bourgeoning life science industry. Theorizing laboratory labor through pragmatist discussion on craftwork, the chapter shows that biomedical research is essentially an embodied craft and the skill of cellular reprogramming begins as bodily practices.
  • Preprint
    Full-text available
    Plagiarism, which Indiana University's Writing Tutorial Service defines as "using others' ideas and words without clearly acknowledging the source of that information", is often described as a major problem. However, standard definitions such as this one suggest only limited solutions to the problem: acknowledging sources or forbidding reuse. Since all scholarship involves engaging with the ideas of others and academic writing tends to reuse certain expressions, these solutions-though important-are of limited utility. This paper examines a type of plagiarism to which the standard solutions do not apply: the reuse of linguistic models without sufficient attention to the logic or thoughts the texts express. We present two cases of plagiarism, from which we can see that plagiarism shows a gap between the written texts and the thoughts of the author. In order to fill the gap, one needs to know how to integrate not only the texts borrowed from others into one's writing but also the thoughts expressed by the texts. Thus a satisfactory solution to the plagiarism problem requires not only writing skills but also logical thinking skills.
  • Article
    Full-text available
    Pluripotent stem cells can be induced from somatic cells, providing an unlimited cell resource, with potential for studying disease and use in regenerative medicine. However, genetic manipulation and technically challenging strategies such as nuclear transfer used in reprogramming limit their clinical applications. Here, we show that pluripotent stem cells can be generated from mouse somatic cells at a frequency up to 0.2% using a combination of seven small-molecule compounds. The chemically induced pluripotent stem cells resemble embryonic stem cells in terms of their gene expression profiles, epigenetic status, and potential for differentiation and germline transmission. By using small molecules, exogenous “master genes” are dispensable for cell fate reprogramming. This chemical reprogramming strategy has potential use in generating functional desirable cell types for clinical applications.
  • Article
    Full-text available
    Multilineage-differentiating stress-enduring (Muse) cells are distinct stem cells in mesenchymal cell populations with the capacity to self-renew, to differentiate into cells representative of all three germ layers from a single cell, and to repair damaged tissues by spontaneous differentiation into tissue-specific cells without forming teratomas. We describe step-by-step procedures for isolating and evaluating these cells. Muse cells are also a practical cell source for human induced pluripotent stem (iPS) cells with markedly high generation efficiency. They can be collected as cells that are double positive for stage-specific embryonic antigen-3 (SSEA-3) and CD105 from commercially available mesenchymal cells, such as adult human bone marrow stromal cells and dermal fibroblasts, or from fresh adult human bone marrow samples. Under both spontaneous and induced differentiation conditions, they show triploblastic differentiation. It takes 4-6 h to collect and 2 weeks to confirm the differentiation and self-renewal capacity of Muse cells.
  • Article
    Previous studies of serial cloning in animals showed a decrease in efficiency over repeated iterations and a failure in all species after a few generations. This limitation led to the suggestion that repeated recloning might be inherently impossible because of the accumulation of lethal genetic or epigenetic abnormalities. However, we have now succeeded in carrying out repeated recloning in the mouse through a somatic cell nuclear transfer method that includes a histone deacetylase inhibitor. The cloning efficiency did not decrease over 25 generations, and, to date, we have obtained more than 500 viable offspring from a single original donor mouse. The reprogramming efficiency also did not increase over repeated rounds of nuclear transfer, and we did not see the accumulation of reprogramming errors or clone-specific abnormalities. Therefore, our results show that repeated iterative recloning is possible and suggest that, with adequately efficient techniques, it may be possible to reclone animals indefinitely.
  • Article
    Full-text available
    Balanced organogenesis requires the orchestration of multiple cellular interactions to create the collective cell behaviours that progressively shape developing tissues. It is currently unclear how individual, localized parts are able to coordinate with each other to develop a whole organ shape. Here we report the dynamic, autonomous formation of the optic cup (retinal primordium) structure from a three-dimensional culture of mouse embryonic stem cell aggregates. Embryonic-stem-cell-derived retinal epithelium spontaneously formed hemispherical epithelial vesicles that became patterned along their proximal-distal axis. Whereas the proximal portion differentiated into mechanically rigid pigment epithelium, the flexible distal portion progressively folded inward to form a shape reminiscent of the embryonic optic cup, exhibited interkinetic nuclear migration and generated stratified neural retinal tissue, as seen in vivo. We demonstrate that optic-cup morphogenesis in this simple cell culture depends on an intrinsic self-organizing program involving stepwise and domain-specific regulation of local epithelial properties.
  • Article
    Two kinds of human pluripotent cells, human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), promise new avenues for medical innovation. These human cells share many similarities with mouse counterparts, including pluripotency, and they exhibit several unique properties. This review examines the diversity of mammalian pluripotent cells from a perspective of metastable pluripotency states. An intriguing phenomenon unique to human pluripotent stem cells is dissociation-induced apoptosis, which has been a technical problem for various cellular manipulations. The discovery that this apoptosis is suppressed by ROCK inhibitors brought revolutionary change to this troublesome situation. We discuss possible links of the metastable pluripotent state to ROCK-dependent human embryonic stem cell apoptosis and summarize recent progress in molecular understandings of this phenomenon.
  • Article
    In female mammals, one of two X chromosomes is epigenetically inactivated for gene dosage compensation, known as X inactivation (Xi). Inactivation occurs randomly in either the paternal or maternal X chromosome in all embryonic cell lineages, designated as random Xi. By contrast, in extra-embryonic cell lineages, which are segregated from somatic cell lineages in pre-implantation development, the paternal X chromosome is selectively inactivated, known as imprinted Xi. Although it is speculated that erasure of the imprinted mark on either the maternal or paternal X chromosome in somatic cell lineages might change the mode of Xi from imprinted to random, it is not known when this event is completed in development. Here, we tested the mode of Xi during the differentiation of female mouse embryonic stem (ES) cells derived from the inner cell mass (ICM) of blastocyst-stage embryos toward trophectoderm (TE) and primitive endoderm (PrE) lineages induced by artificial activation of transcription factor genes Cdx2 and Gata6, respectively. We found that random Xi occurs in both TE and PrE cells. Moreover, cloned embryos generated by the transfer of nuclei from the female ES cells showed random Xi in TE, suggesting the complete erasure of all X imprints for imprinted Xi in ICM-derived ES cells.
  • Article
    Full-text available
    Mature adult tissues contain stem cells that express many genes normally associated with the early stage of embryonic development, when maintained in appropriate environments. Cells procured from adult tissues representative of the three germ layers (spinal cord, muscle, and lung), each exhibiting the potential to mature into cells representative of all three germ layers. Cells isolated from adult tissues of different germ layer origin were propagated as nonadherent clusters or spheres that were composed of heterogeneous populations of cells. When the clusters or spheres were dissociated, the cells had the ability to reform new, nonadherent spheres for several generations. When implanted in vivo, in association with biodegradable scaffolds, into immunodeficient mice, tissue containing cells characteristic of the three germ layers was generated. These findings suggest the existence of a population of stem cells in adult tissues that is quite different and distinct from embryonic stem cells that demonstrate a greater potency for differentiation across germ lines than previously believed. Such cells could potentially be as useful as embryonic stem cells in tissue engineering and regenerative medicine.
  • Article
    Human embryonic stem cells (hESCs), unlike mouse ones (mESCs), are vulnerable to apoptosis upon dissociation. Here, we show that the apoptosis, which is of a nonanoikis type, is caused by ROCK-dependent hyperactivation of actomyosin and efficiently suppressed by the myosin inhibitor Blebbistatin. The actomyosin hyperactivation is triggered by the loss of E-cadherin-dependent intercellular contact and also observed in dissociated mouse epiblast-derived pluripotent cells but not in mESCs. We reveal that Abr, a unique Rho-GEF family factor containing a functional Rac-GAP domain, is an indispensable upstream regulator of the apoptosis and ROCK/myosin hyperactivation. Rho activation coupled with Rac inhibition is induced in hESCs upon dissociation, but not in Abr-depleted hESCs or mESCs. Furthermore, artificial Rho or ROCK activation with Rac inhibition restores the vulnerability of Abr-depleted hESCs to dissociation-induced apoptosis. Thus, the Abr-dependent "Rho-high/Rac-low" state plays a decisive role in initiating the dissociation-induced actomyosin hyperactivation and apoptosis in hESCs.
  • Article
    During early mammalian development, as the pluripotent cells that give rise to all of the tissues of the body proliferate and expand in number, they pass through transition states marked by a stepwise restriction in developmental potential and by changes in the expression of key regulatory genes. Recent findings show that cultured stem-cell lines derived from different stages of mouse development can mimic these transition states. They further reveal that there is a high degree of heterogeneity and plasticity in pluripotent populations in vitro and that these properties are modulated by extrinsic signalling. Understanding the extrinsic control of plasticity will guide efforts to use human pluripotent stem cells in research and therapy.
  • Article
    Full-text available
    We found adult human stem cells that can generate, from a single cell, cells with the characteristics of the three germ layers. The cells are stress-tolerant and can be isolated from cultured skin fibroblasts or bone marrow stromal cells, or directly from bone marrow aspirates. These cells can self-renew; form characteristic cell clusters in suspension culture that express a set of genes associated with pluripotency; and can differentiate into endodermal, ectodermal, and mesodermal cells both in vitro and in vivo. When transplanted into immunodeficient mice by local or i.v. injection, the cells integrated into damaged skin, muscle, or liver and differentiated into cytokeratin 14-, dystrophin-, or albumin-positive cells in the respective tissues. Furthermore, they can be efficiently isolated as SSEA-3(+) cells. Unlike authentic ES cells, their proliferation activity is not very high and they do not form teratomas in immunodeficient mouse testes. Thus, nontumorigenic stem cells with the ability to generate the multiple cell types of the three germ layers can be obtained through easily accessible adult human mesenchymal cells without introducing exogenous genes. These unique cells will be beneficial for cell-based therapy and biomedical research.