Embryonic-like Stem Cells Research Articles

A novel investigation of NANOG and POU5F1 associations in the pluripotent characterization of ES-like and epiblast cells

The transcription factors NANOG and POU5F1 (OCT4) play crucial roles in maintaining pluripotency in embryonic stem (ES) cells. While their functions have been well-studied, the specific interactions between NANOG and POU5F1 and their combined effects on pluripotency in ES-like and Epiblast cells remain less understood. Understanding these associations is vital for refining pluripotent stem cell characterization and advancing regenerative medicine. In this matter, we investigated the associations between NANOG and POU5F1 in maintaining pluripotency in ES-like and Epiblast cells and how these interactions contribute to the distinct pluripotent states of these cells. In the present paper, we examined the pattern of NANOG expression by the immunocytochemical method in embryonic stem-like (ES-like) cells and compared it with its expression pattern in embryonic stem cells (ESCs). Similarly, we examined the expression pattern of POU5F1 in ES-like cells, ESCs, and epiblast cells and compared the expression pattern of these two genes with each other. On the other hand, using Fluidigm Biomark system analysis, we compared the amount of NANOG mRNA in these three cell lines and differentiated and undifferentiated Spermatogonial stem cells in several passages. Microscopic observations indicated the cytoplasmic expression of NANOG in the considered cells; moreover, they showed a similar expression pattern of NANOG with POU5F1 in the experimented cells. It has also been suggested that the more limited the cell’s pluripotency, the lower the expression of these two genes. However, the decrease in NANOG expression is less than that of POU5F1. Fluidigm real-time RT-PCR analysis also confirmed these results. During the experimental process, protein-protein (PPI) network analysis shows a significant association of NANOG with other stem cell proteins, such as POU5F1. Our findings reveal distinct yet overlapping roles of NANOG and POU5F1 in maintaining pluripotency in ES-like and Epiblast cells. The differential binding patterns and functional interactions between these factors underscore the complexity of pluripotency regulation in different stem cell states. This study provides new insights into the molecular mechanisms governing pluripotency and highlights potential targets for enhancing stem cell-based therapies.

Selected stem cell populations in pediatric acute lymphoblastic leukemia.

Acute lymphoblastic leukemia is characterized by a disturbed maturation of hematopoietic stem cells (HSCs) resulting in development of a malignant clone. Despite relatively positive outcome, there are still instances of disease relapse occurring due to ineffective disease eradication or primary leukemic clone alterations. Unclear significance of stem cells in the course of ALL led us to investigate and establish crucial changes in two stem cell populations - very small embryonic-like stem cells (VSELs) and HSCs during the induction phase of treatment. In a retrospective study selected stem cells in peripheral blood and bone marrow of 60 pediatric ALL subjects and 48 healthy controls were subjected to flow cytometric analysis at 4 different time points. Both VSELs and HSCs were elevated at the moment of ALL diagnosis compared to healthy controls, but profoundly decline until day 15. Further observations revealed an increase in HSCs with a concomitant depletion of VSELs until week 12. ALL patients with high HSCs showed positive correlation with bone marrow blasts at diagnosis. Patients with lower VSELs or HSCs at diagnosis had slightly improved response to applied therapy. We observed higher initial bone marrow lymphoblast values in patients with lower VSELs or higher HSCs in the high-risk group. The significance of VSELs in predicting treatment outcome can be illustrated by lower day 15 MRD level of patients with lower VSELs at diagnosis. We found HSCs and VSELs to be valid participants in pediatric ALL with possible contribution in the neoplastic process and prediction of initial treatment outcome.

Unraveling the Significance of Nanog in the Generation of Embryonic Stem-like Cells from Spermatogonia Stem Cells: A Combined In Silico Analysis and In Vitro Experimental Approach.

Embryonic stem-like cells (ES-like cells) are promising for medical research and clinical applications. Traditional methods involve "Yamanaka" transcription (OSKM) to derive these cells from somatic cells in vitro. Recently, a novel approach has emerged, obtaining ES-like cells from spermatogonia stem cells (SSCs) in a time-related process without adding artificial additives to cell cultures, like transcription factors or small molecules such as pten or p53 inhibitors. This study aims to investigate the role of the Nanog in the conversion of SSCs to pluripotent stem cells through both in silico analysis and in vitro experiments. We used bioinformatic methods and microarray data to find significant genes connected to this derivation path, to construct PPI networks, using enrichment analysis, and to construct miRNA-lncRNA networks, as well as in vitro experiments, immunostaining, and Fluidigm qPCR analysis to connect the dots of Nanog significance. We concluded that Nanog is one of the most crucial differentially expressed genes during SSC conversion, collaborating with critical regulators such as Sox2, Dazl, Pou5f1, Dnmt3, and Cdh1. This intricate protein network positions Nanog as a pivotal factor in pathway enrichment for generating ES-like cells, including Wnt signaling, focal adhesion, and PI3K-Akt-mTOR signaling. Nanog expression is presumed to play a vital role in deriving ES-like cells from SSCs in vitro. Finding its pivotal role in this path illuminates future research and clinical applications.

Initiation of Cancer: The Journey From Mutations in Somatic Cells to Epigenetic Changes in Tissue-resident VSELs.

Multiple theories exist to explain cancer initiation, although a consensus on this is crucial for developing effective therapies. 'Somatic mutation theory' suggests that mutations in somatic cells during DNA repair initiates cancer but this concept has several attached paradoxes. Research efforts to identify quiescent cancer stem cells (CSCs) that survive therapy and result in metastasis and recurrence have remained futile. In solid cancers, CSCs are suggested to appear during epithelial-mesenchymal transition by the dedifferentiation and reprogramming of epithelial cells. Pluripotent and quiescent very small embryonic-like stem cells (VSELs) exist in multiple tissues but remain elusive owing to their small size and scarce nature. VSELs are developmentally connected to primordial germ cells, undergo rare, asymmetrical cell divisions and are responsible for the regular turnover of cellsto maintain tissuehomeostasis throughout life. VSELs are directly vulnerable to extrinsic endocrine insults because they express gonadal and gonadotropin hormone receptors. VSELs undergo epigenetic changes due to endocrine insults and transform into CSCs. CSCs exhibit genomic instability and develop mutations due to errors during DNA replication while undergoing excessive proliferation and clonal expansion to formspheroids. Thus tissue-resident VSELs offer a connection between extrinsic insults and variations in cancer incidence reported in various body tissues. To conclude, cancer is indeed a stem cell disease with mutations occurring as a consequence. In addition to immunotherapy, targeting mutations, and Lgr5 + organoids for developing new therapeutics, targeting CSCs (epigenetically altered VSELs) by improving their niche and epigenetic status could serve as a promising strategy to treat cancer.

Identification and quantification of very small embryonic-like (VSEL) stem cells in fresh umbilical cord blood

The present study aimed to identify, quantify and compare the quantities of very small embryonic-like (VSEL) stem cells in the two main fractions of fresh umbilical cord blood (UCB). To do this, UCB obtained during caesarean section underwent gradient centrifugation. Nucleated cells isolated from the two main fractions – the mononuclear cell layer above the gradient, and the red blood cell pellet below the gradient – were immunofluorescently labelled to identify the VSELs (CD45−/CD133+/SSEA4+). These two cell fractions were then analysed via multiparameter sorting on a flow cytometer and the quantities of positively stained cells were compared.

Metadichol, a Natural Ligand for the Expression of Yamanaka Reprogramming Factors in Human Cardiac, Fibroblast, and Cancer Cell Lines

The reprogramming of somatic cells into embryonic-like stem cells requires the activation of four essential transcription factors, referred to as the Yamanaka factors: Sox2, Oct4, Klf4, and c-Myc. However, the utilization of these genes and viral vectors for their delivery presents a risk of carcinogenesis, which renders induced pluripotent stem cell (iPSC) lines unsuitable for clinical applications. Typically, direct reprogramming involves the use of viral vectors to induce the expression of these factors. In contrast, metadichol, a novel approach, enhances the expression of Yamanaka factors in cells using a dose ranging from 1 pg to 100 ng, thereby preventing the need for viral vectors. This non-viral method renders cellular reprogramming safer and more clinically viable. Treatment with metadichol within the specific dose range resulted in a substantial augmentation of the fibroblast expression of OCT4, SOX2, KLF4, KLF2, and Nanog, which was confirmed through qRT-PCR and western blot analyses. The expression levels of OCT4, KLF4, Nanog, and Sox2, exhibited an increase of 4.01-, 3.51-, 1.26-, and 2.5-fold, respectively, compared to the controls. Notably, A549 and Colo-205 cancer cells demonstrated a marked elevation in expression levels. The reprogramming of primary human cancer cells presents considerable challenges. However, in triple-negative primary breast cancer cells, metadichol treatment led to the substantial upregulation of the expression levels of OCT4, KLF4, Nanog, and Sox2 by 19.6-, 8.07-, 2.45-, and 6.91-fold, respectively, across the dose range of 1 pg to 100 ng. Klotho, an antiaging gene that is modulated by metadichol, down regulates TP53, a crucial factor for the production of somatic cell iPSCs. Additionally, metadichol enhances the availability of vitamin C, which is essential for generating iPSCs from somatic cells. Increased levels of alkaline phosphatase, indicative of cellular differentiation, were observed in the treated group compared to the controls. Metadichol is a nontoxic nanoemulsion that is derived from long-chain C26–C28 alcohols from food sources and activates Yamanaka factors across various cell types without requiring the use of viral or CRISPR-based methodologies, thus substantially advancing somatic cell reprogramming.

(Very) Small Stem-like Cells in Human Cell Cultures

Very Small Embryonic-like Stem Cells (VSELSCs) and Very Small Cancer Stem Cells (VSCSCs) are fields of intensive research. Although the presence in vitro of VSELSC and VSCSC cellular stage analogs appear probable, it has yet to be published. Utilizing established human cell cultures with varying populations of primitive cells, stained with CD markers specific to primitive stages, in addition to a fluorescent DNA dye, and following histochemical processing, we have developed a cytological method for detecting Very Small Leukemic Stem-like Cells (VSLSLCs), Very Small Cancer Stem-like Cells (VSCSLCs), and VSELSCs. This detection provides an opportunity to advance research in these areas.

Reversing Uteropathies Including Cancer-Like Changes in Mice by Transplanting Mesenchymal Stromal Cells or XAR Treatment.

Pluripotent, very small embryonic-like stem cells (VSELs) and tissue-committed 'progenitors' termed endometrial stem cells (EnSCs) are reported in mouse uterus. They express gonadal and gonadotropin hormone receptors and thus are vulnerable to early-life endocrine insults. Neonatal exposure of mouse pups to endocrine disruption cause stem/progenitor cells to undergo epigenetic changes, excessive self-renewal, and blocked differentiation that results in various uteropathies including non-receptive endometrium, hyperplasia, endometriosis, adenomyosis, and cancer-like changes in adult life. Present study investigated reversal of these uteropathies, by normalizing functions of VSELs and EnSCs. Two strategies were evaluated including (i) transplanting mesenchymal stromal cells (provide paracrine support) on D60 or (ii) oral administration of XAR (epigenetic regulator) daily from days 60-100 and effects were studied later in 100 days old mice. Results show normalization of stem/progenitor cells (Oct-4, Oct-4A, Sox-2, Nanog) and Wnt signalling (Wnt-4, β-catenin, Axin-2) specific transcripts. Flow cytometry results showed reduced numbers of 2-6 µm, LIN-CD45-SCA-1 + VSELs. Hyperplasia (Ki67) of epithelial (Pax-8, Foxa-2) and myometrial (α-Sma, Tgf-β) cells was reduced, adenogenesis (differentiation of glands) was restored, endometrial receptivity and differentiation (LIF, c-KIT, SOX-9, NUMB) and stromal cells niche (CD90, VIMENTIN, Pdgfra, Vimentin) were improved, cancer stem cells markers (OCT-4, CD166) were reduced while tumor suppressor genes (PTEN, P53) and epigenetic regulators (Ezh-2, Sirt-1) were increased. To conclude, normalizing VSELs/EnSCs to manage uteropathies provides a novel basis for initiating clinical studies. The study falls under the umbrella of United Nations Sustainable Development Goal 3 to ensure healthy lives and well-being for all of all ages.

Current Progress in Stem Cell Therapy for Male Infertility.

Infertility has become one of the most common issues worldwide, which has negatively affected society and infertile couples. Meanwhile, male infertility is responsible for about 50% of infertility. Accordingly, a great number of researchers have focused on its treatment during the last few years; however, current therapies such as assisted reproductive technology (ART) are not effective enough in treating male infertility. Because of their self-renewal and differentiation capabilities and unlimited sources, stem cells have recently raised great hope in the treatment of reproductive system disorders. Stem cells are undifferentiated cells that can induce different numbers of specific cells, such as male and female gametes, demonstrating their potential application in the treatment of infertility. The present review aimed at identifying the causes and potential factors that influence male fertility. Besides, we highlighted the recent studies that investigated the efficiency of stem cells such as spermatogonial stem cells (SSCs), embryonic stem cells (ESCs), very small embryonic-like stem cells (VSELs), induced pluripotent stem cells (iPSCs), and mesenchymal stem cells (MSCs) in the treatment of various types of male infertility.

Endometrial and placental stem cells in successful and pathological pregnancies.

The endometrium is a dynamic tissue that undergoes extensive remodeling during the menstrual cycle and further gets modified during pregnancy. Different kinds of stem cells are reported in the endometrium. These include epithelial stem cells, endometrial mesenchymal stem cells, side population stem cells, and very small embryonic-like stem cells. Stem cells are also reported in the placenta which includes trophoblast stem cells, side population trophoblast stem cells, and placental mesenchymal stem cells. The endometrial and placental stem cells play a pivotal role in endometrial remodeling and placental vasculogenesis during pregnancy. The dysregulation of stem cell function is reported in various pregnancy complications like preeclampsia, fetal growth restriction, and preterm birth. However, the mechanisms by which it does so are yet elusive. Herein, we review the current knowledge of the different type of stem cells involved in pregnancy initiation and also highlight how their improper functionality leads to pathological pregnancy.

Molecular analysis and comparison of CD34+ and CD133+ very small embryonic-like stem cells purified from umbilical cord blood.

Very small embryonic like stem cells (VSELs) are a dormant population of stem cells that, as proposed, are deposited during embryogenesis in various tissues, including bone marrow (BM). These cells are released under steady state conditions from their tissue locations and circulate at a low level in peripheral blood (PB). Their number increases in response to stressors as well as tissue/organ damage. This increase is evident during neonatal delivery, as delivery stress prompts enrichment of umbilical cord blood (UCB) with VSELs. These cells could be purified from BM, PB, and UCB by multiparameter sorting as a population of very small CXCR4+ Lin- CD45- cells that express the CD34 or CD133 antigen. In this report, we evaluated a number of CD34+ Lin- CD45- and CD133+ Lin- CD45- UCB-derived VSELs. We also performed initial molecular characterization of both cell populations for expression of selected pluripotency markers and compared these cells at the proteomic level. We noticed that CD133+ Lin- CD45- population is more rare and express, at a higher level, mRNA for pluripotency markers Oct-4 and Nanog as well as the stromal-derived factor-1 (SDF-1) CXCR4 receptor that regulates trafficking of these cells, however both cells population did not significantly differ in the expression of proteins assigned to main biological processes.

Candidates for early detection: Effect of pluripotent very small embryonic-like stem cells (VSELs) to transform into cancer stem cells (CSCs) and candidacy for early detection of cancer in a liquid biopsy.

e15024 Background: Prevailing somatic mutation theory (SMT) is mired with several paradoxes and still there is no clarity on how cancer initiates. No distinct pattern of somatic driver mutations specific to cancers affecting solid tissues has yet emerged even after massive NGS studies since 2007. Rather than cancer being a genetic disease, we postulate that endogenous, tissue-resident VSELs get transformed into CSCs to initiate cancer. Under normal conditions, VSELs along with tissue-specific progenitors work in a subtle manner to maintain life-long homeostasis in adult tissues. VSELs express steroid and gonadotropin hormone receptors, are directly vulnerable to environmental insults, and their dysfunction initiates various pathologies, including cancer. Recent mice studies have shown that environmental insults resulted in global hypomethylation, genomic instability and loss of imprinting (LOI) at IGF2/H19 loci that transformed quiescent VSELs into CSCs. Unlike VSELs, CSCs undergo excessive self-renewal, acquire bad-luck mutations during clonal expansion and oncogene activation to initiate cancers, and can also further mobilize to distant sites to cause metastasis. A 1000-sample clinical study based on OCT-4A expression by qRT-PCR suggested that CSCs that get mobilized from affected organs can serve as potential novel candidates for early detection of cancer through liquid biopsy. Methods: CSCs were enriched from the peripheral blood using our proprietary technology. They were enumerated by flow cytometry. VSELs undergo epigenetic changes and LOI to transform into CSCs. TaqMan based qRT-PCR was carried out to study OCT-4A and ALDH-1 positive CSCs and IGF2 expression to monitor LOI at the IGF2/H19 loci. LOI of IGF2 is commonly associated with initiation and also progression of multiple cancers and is also responsible for pushing VSELs out of quiescence and enter the cell cycle as CSCs. Results: Increased numbers of CSCs (epigenetically altered VSELs) were detected in peripheral blood samples of cancer subjects compared to normal subjects (p < 0.0001) by flow cytometry studies. The median absolute count of circulatory VSELs/CSCs was 190 per ml (IQR: 105-295) in normal subjects and 690 per ml (IQR: 370-1265) in cancer subjects. The qRT-PCR study conducted on 100 subjects showed increased expression of OCT-4A, IGF2 and ALDH1 genes in cancer compared to non-cancer subjects. Conclusions: Early detection of cancer by evaluating circulating tumor cells (CTCs) or tumor DNA (ctDNA) holds promise, but their scarce nature and mobilization in late stages only when tumor is > 3 mm in size, are major limitations for widespread translation. Circulating VSELs/CSCs in peripheral blood are evidently promising candidates to screen and classify subjects for cancer absence, likely risk, or presence.

Transformation of tissue-resident, very small embryonic-like stem cells (VSELs) into cancer stem cells (CSCs) and its role in cancer initiation via epigenetic changes.

e15084 Background: CSCs are postulated to initiate cancer but still no consensus on their surface markers and origin. Pluripotent VSELs exist in adult tissues, express receptors for steroid and gonadotropin hormones and are possible ‘embryonic remnants’ that could initiate cancer. Methods: Neonatal exposure to diethylstilbestrol (DES) or estradiol (E2) resulted in tumor-like changes in uterus and testes of 100 days old mice. Various studies included histological changes, VSELs enumeration by flow cytometry, and a carefully selected panel of transcripts by qRT-PCR to delineate various cellular events. Mice neonatally exposed to endocrine disruption were administered XAR (a nano-formulation of resveratrol) or transplanted niche-providing mesenchymal stromal cells on D60 and studied later on D100 for reversal/prevention of cancer phenotype. Results: Both E2 and DES resulted in increased numbers of VSELs in testis (Ctrl:0.2%; E2:0.4%; DES:1.3%) and uterus (Ctrl:18.52%; E2:35.42%; DES:40.57%). An up-regulation of pluripotent transcripts ( Oct-4, Sox-2, Nanog) was detected. Oct4A (specific marker for pluripotent VSELs) was upregulated after both DES (testis > 8 folds; uterus > 15 folds) and E2 (testis > 3 folds and uterus > 10 folds) treatment. Excessive self-renewal of VSELs was associated with dysregulated expression of DNA methyltransferases (Dnmts). Dnmt1 was downregulated while Dnmt3a, Dnmt3b, and Dnmt3L were upregulated in the testis. In the uterus, Dnmts showed increased expression after both E2 and DES treatment. Marked reduction of 5mC suggested global hypomethylation and increased Igf2 and Dlk-1 suggested affected/loss of imprinting at IGF2/H19 and DLK-1/MEG3 loci. VSELs acquired CSCs phenotype with increased expression of CD166 and ALDH1. Genomic instability in CSCs was suggested by dysregulated mismatch repair axis (DNMT-1/NP95/PCNA) and repair enzymes ( Brca-1, Rad-51, Mlh-1). Reduced expression of Pten and p53 was associated with blocked differentiation. Both reversal strategies resulted in normalization of stem cells compartment, improved differentiation (spermatogenesis/adenogenesis), reduced CSC markers, and increased p53, Pten, Np95, Sirt-1 expression. Interestingly, XAR treatment resulted in downregulation of Igf2 and Dlk1. Stem cells niche was also improved as evident by increased expression of SOX9 in testis and CD90, VIMENTIN in endometrium. Conclusions: Only immortal VSELs can transmit developmental insults to adult cells. Altered epigenetic state of VSELs resulted in cancer initiation and it was possible to prevent/reverse cancer by normalizing epigenetic state of VSELs and their niche. Targeting CSCs and pushing them back into quiescent VSELs and facilitating their normal differentiation offers novel method to win the war against cancer.

Beyond brain injury biomarkers: chemoattractants and circulating progenitor cells as biomarkers of endogenous rehabilitation effort in preterm neonates with encephalopathy.

Preclinical work and studies in adults have shown that endogenous regeneration efforts that involve mobilization of progenitor cells take place after brain injury. However, kinetics of endogenous circulating progenitor cells (CPCs) in preterm neonates is not well described, particularly their possible role regarding brain injury and regeneration. We aimed to assess the kinetics of CPCs in neonates with encephalopathy of prematurity in relation to brain injury biomarkers, chemoattractants and relevant antenatal and postanal clinical factors, in an effort to outline the related pathophysiology. 47 preterm neonates (of 28-33 weeks GA) were enrolled: 31 newborns with no or minimal brain injury (grade I IVH) and 16 prematures with encephalopathy (grade III or IV IVH, PVL or infarct). Peripheral blood samples obtained on days 1, 3, 9, 18 and 45 after birth were analyzed using flow cytometry, focusing on EPCs (early and late Endothelial Progenitor Cells), HSCs (Hematopoietic Stem Cells) and VSELs (Very Small Embryonic-Like Stem Cells). At the same time-points serum levels of S100B, Neuron-specific Enolase (NSE), Erythropoietin (EPO), Insulin-like growth factor-1 (IGF-1) and SDF-1 were also measured. Neonates were assessed postnatally with brain MRI, and with Bayley III developmental test at 2 years of corrected age. Preterms with brain injury proved to have significant increase of S100B and NSE, followed by increase of EPO and enhanced mobilization mainly of HSCs, eEPCs and lEPCs. IGF-1 was rather decreased in this group of neonates. IGF-1 and most CPCs were intense decreased in cases of antenatal or postnatal inflammation. S100B and NSE correlated with neuroimaging and language scale in Bayley III test, providing good prognostic ability. The observed pattern of CPCs' mobilization and its association with neurotrophic factors following preterm brain injury indicate the existence of an endogenous brain regeneration process. Kinetics of different biomarkers and associations with clinical factors contribute to the understanding of the related pathophysiology and might help to early discriminate neonates with adverse outcome. Timely appropriate enhancement of the endogenous regeneration effort, when it is suppressed and insufficient, using neurotrophic factors and exogenous progenitor cells might be a powerful therapeutic strategy in the future to restore brain damage and improve the neurodevelopmental outcome in premature infants with brain injury.

Very Small Embryonic-Like Stem Cells

Continuous and growing research studies regarding the clinical applications of the pluripotent or multipo-tent stem cells with their potential to differentiate into three germ layers are very well conducted in regenerative medicine (RM). In this review, we report the recent clinical applications and potential use of very small embryonic-like stem cells (VSELs) in orthopedics. VSELs are nonhematopoietic (CD45 - / Lin -), rare, and very small cells; they were reported as “dormant” cells in the bone marrow (BM), but are also found in cord blood, peripheral blood (PB), and in adult organs. Based on their capability to express markers of pluripotency (such as Oct-4 +/Nanog +/SSEA-1/4+/CXCR4+), it has been hypothesized that these cells could be early deposited during the embryonic development as descendants of epiblast-derived stem cells and perhaps from some primordial germ cells. VSELs can be released or mobilized from the BM to the PB during tissue injury and stress, facilitating the regeneration of damaged tissues. As well as mesenchymal stem cells, nowadays VSELs can be expanded ex vivo. Their pluripotency could be suitable for applications in RM, solving several problems regarding the use of both controversial embryonic stem cells and induced pluripotent stem cells. VSELs studies will hopefully open new frontiers to better understand their potential that would be relevant for future applications in RM and translational research.

Retraction of: Detection, Characterization, and Spontaneous Differentiation In Vitro of Very Small Embryonic-Like Putative Stem Cells in Adult Mammalian Ovary (10.1089/scd.2010.0461).

Stem Cells and DevelopmentAhead of Print Retraction NoticeFree AccessRetraction of: Detection, Characterization, and Spontaneous Differentiation In Vitro of Very Small Embryonic-Like Putative Stem Cells in Adult Mammalian Ovary (10.1089/scd.2010.0461)This article retracts the following:Retraction: Detection, Characterization, and Spontaneous Differentiation In Vitro of Very Small Embryonic-Like Putative Stem Cells in Adult Mammalian OvaryPublished Online:6 May 2023https://doi.org/10.1089/scd.2010.0461.retractAboutSectionsPDF/EPUB Permissions & CitationsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail The Editor-in-Chief of Stem Cells and Development officially retracts the article entitled, “Detection, Characterization, and Spontaneous Differentiation In Vitro of Very Small Embryonic-Like Putative Stem Cells in Adult Mammalian Ovary,” by Seema Parte, Deepa Bhartiya, Jyoti Telang, Vinita Daithankar, Vinita Salvi, Kusum Zaveri, and Indira Hinduja (Stem Cells Dev 2011;20(8):1451-1464; doi: 10.1089/scd.2010.0461).A reported comment on the PubPeer platform suggested that the “images in Figure 6D and Figure 6F appear to overlap but are described differently. The images are oriented differently, have different contrast, and are at a different magnification (red boxes are not the same size). Difficult to understand how these transformations would occur merely as errors during figure assembly,” (PubPeer, 2022).The Editor-in-Chief of the journal contacted the corresponding author of the paper, Dr. Deepa Bhartiya, requesting a “prompt response and intended course of action” to the questions raised on PubPeer. Dr. Bhartiya responded, indicating that the errors in both figures were “an unintentional human error.”After a series of communications between Dr. Bhartiya and the Editor-in-Chief of Stem Cells and Development, Dr. Bhartiya claimed full responsibility for the contents of the article and requested that corrections to the paper by removing the incorrect images, as indicated in the email below: A discrepancy in two figures (Fig 6D and negative control in Fig 11) was picked up by an Artificial Intelligence based tool and pointed out on Pubpeer. A careful examination shows that indeed there is a mistake but it was inadvertent and simply an oversight and not a deliberate manipulation of any kind. It is the same figure of a neuron turned up side down and pasted at two places. This remained unnoticed by the authors and also was not picked up by the reviewers or the readers [for] over a decade. It is our humble request to all those who have cited our work to be rest assured that the work integrity is intact. We apologize for the inconvenience caused to those directly and indirectly affected by this process. My students worked with great dedication to acquire the data and obtained results to compile into a manuscript. There is no manipulation whatsoever and this unintentional error remained unnoticed by everyone involved with the manuscript preparation and publication. We will try to submit the revised version again with due permissions. We request the editor to allow us to publish an addendum/erratum to take care of the mistake otherwise it is his decision to retract the paper.” [sicThe Editor of the journal rejected the contention that the manipulation and combining of the same images within the panels purported to represent different experiments could be unintentional, and hence has denied the request to publish an erratum and offered the authors the opportunity to self-retract the article. The authors declined to do so and continued to request a corrigendum. The Editor determined that an editorial retraction was warranted based on the discovery of the discrepancies in the images. Dr. Bhartiya was notified via email of the decision to editorially retract the paper.ReferenceActinopolyspora biskrensis. PubPeer. December 2022. https://pubpeer.com/publications/B68E0F1096533B72D2B37075C258BC?utm_source=Firefox&utm_medium=BrowserExtension&utm_campaign=Firefox Google ScholarFiguresReferencesRelatedDetailsRelated articlesRetraction: Detection, Characterization, and Spontaneous Differentiation In Vitro of Very Small Embryonic-Like Putative Stem Cells in Adult Mammalian Ovary3 Feb 2011Stem Cells and Development Volume 0Issue 0 InformationCopyright 2023, Mary Ann Liebert, Inc., publishersTo cite this article:Retraction of: Detection, Characterization, and Spontaneous Differentiation In Vitro of Very Small Embryonic-Like Putative Stem Cells in Adult Mammalian Ovary (10.1089/scd.2010.0461).Stem Cells and Development.ahead of printhttp://doi.org/10.1089/scd.2010.0461.retractOnline Ahead of Print:May 6, 2023PDF download

Review on the Method of Generating Vascular Cells from iPSCs and Its Application

Induced Pluripotent Stem Cells (iPSCs) embryonic-like stem cells are reprogrammed from skin or adult somatic cells. iPSC-derived cardiovascular cells have been widely used in many cardiovascular disease models and drug discovery. This paper reviews and summarizes the method of generating three different types of iPSC-derived cardiovascular cells, including iPSC-derived Endothelial Cells (ECs), Cardiomyocytes (CMs), and Vascular Smooth Muscle Cells (vSMCs), and the application of these iPSC-derived cardiovascular cells. The diseases discussed in this paper are pulmonary arterial hypertension (PAH) and critical limb ischemia (CLI). Additionally, construction of tissue-engineered blood vessels (TEBVs) for vascular grafting has also been discussed. To conclude, iPSCs, as a promising technique, is used in disease modeling, drug screening, tissue engineering, and the discovery of specific cardiovascular therapies for various patients.

The evolutionary cancer gene network theory versus embryogenic hypotheses.

The present paper compares the statements of the evolutionary cancer gene-network theory (cancer genome theory) with the statements of embryogenic hypotheses like the embryonic rest hypothesis, the very small embryonic-like stem cells hypothesis, (VSEL), the para-embryonic p-ESC hypothesis, and the PGCC-life cycle hypothesis, including the life code theory. In my opinion, the evolutionary gene network theory is the only theory that can satisfactorily explain the homologies between carcinogenesis, tumorigenesis, metastasis, gametogenesis, and early embryogenesis. From an evolutionary point of view, there is no reason to see the origin of cancer in cells of early embryonic life.

Very Small Embryonic-Like Stem Cells Transform Into Cancer Stem Cells and are Novel Candidates for Detecting/Monitoring Cancer by a Simple Blood Test.

Cancer continues to remain a "Black Box" as there is no consensus on how it initiates, progresses, metastasizes, or recurs. Many imponderables exist about whether somatic mutations initiate cancer, do cancer stem cells (CSCs) exist, and if yes, are they a result of de-differentiation or originate from tissue-resident stem cells; why do cancer cells express embryonic markers, and what leads to metastasis and recurrence. Currently, the detection of multiple solid cancers through liquid biopsy is based on circulating tumor cells (CTCs) or clusters, or circulating tumor DNA (ctDNA). However, quantity of starting material is usually adequate only when the tumor has grown beyond a certain size. We posit that pluripotent, endogenous, tissue-resident, very small embryonic-like stem cells (VSELs) that exist in small numbers in all adult tissues, exit from their quiescent state due to epigenetic changes in response to various insults and transform into CSCs to initiate cancer. VSELs and CSCs share properties like quiescence, pluripotency, self-renewal, immortality, plasticity, enrichment in side-population, mobilization, and resistance to oncotherapy. HrC test, developed by Epigeneres, offers the potential for early detection of cancer using a common set of VSEL/CSC specific bio-markers in peripheral blood. In addition, NGS studies on VSELs/CSCs/tissue-specific progenitors using the All Organ Biopsy (AOB) test provide exomic and transcriptomic information regarding impacted organ(s), cancer type/subtype, germline/somatic mutations, altered gene expressions, and dysregulated pathways. To conclude, HrC and AOB tests can confirm the absence of cancer and categorize the rest of subjects into low/moderate/high risk of cancer, and also monitor response to therapy, remission, and recurrence.

A Simplified and Effective Approach for the Isolation of Small Pluripotent Stem Cells Derived from Human Peripheral Blood

Pluripotent stem cells are key players in regenerative medicine. Embryonic pluripotent stem cells, despite their significant advantages, are associated with limitations such as their inadequate availability and the ethical dilemmas in their isolation and clinical use. The discovery of very small embryonic-like (VSEL) stem cells addressed the aforementioned limitations, but their isolation technique remains a challenge due to their small cell size and their efficiency in isolation. Here, we report a simplified and effective approach for the isolation of small pluripotent stem cells derived from human peripheral blood. Our approach results in a high yield of small blood stem cell (SBSC) population, which expresses pluripotent embryonic markers (e.g., Nanog, SSEA-3) and the Yamanaka factors. Further, a fraction of SBSCs also co-express hematopoietic markers (e.g., CD45 and CD90) and/or mesenchymal markers (e.g., CD29, CD105 and PTH1R), suggesting a mixed stem cell population. Finally, quantitative proteomic profiling reveals that SBSCs contain various stem cell markers (CD9, ITGA6, MAPK1, MTHFD1, STAT3, HSPB1, HSPA4), and Transcription reg complex factors (e.g., STAT5B, PDLIM1, ANXA2, ATF6, CAMK1). In conclusion, we present a novel, simplified and effective isolating process that yields an abundant population of small-sized cells with characteristics of pluripotency from human peripheral blood.