Embryonic-like Stem Cells Research Articles

Blood Traffic of Endothelial, Mesenchymal, Hematopoietic and Lin-/CD45-CD34+AC133+CXCR4+ Progenitor Cell Subsets Show Different Patterns in Pre-Fibrotic and Fibrotic Primary Myelofibrosis

Introduction. Primary myelofibrosis (PMF) can be associated to increased bloodstream traffic of some adult progenitor cells. The extramedullary hematopoiesis developed in this setting might remind some features of the fetal hematopoiesis.Material and methods. Some evidence of this hypothesis was searched by flow-cytometry and molecular analysis of blood samples from 12 untreated patients with PMF, 6 pre-fibrotic (PF) and 6 fibrotic (F). The PF or F phase was defined according to the WHO and Thiele histopathology scores.Results. As expected we detected high numbers of CD34+ cells (258x103±402x103/ml; range 1.2-1218x103/ml) as well as small mean numbers of Lin-/CD45-CD34+AC133+CXCR4+ progenitor cells (130±275/ml; range 0-882/ml) that are related to VSELs (very-small embryonic-like stem cells), which have been described as putative pluripotent adult stem cells. When we looked at some embryonic molecular markers by RT-PCR, NANOG and OCT4 expression was detected in the MNC fraction of all the patients tested (hES and CPRE2 cell lines were the positive and negative controls). OCT4 expression was half the level of hES being stronger in F-PMF than in PF-PMF. In all the tested patients NANOG expression was similar to that of hES, whereas SOX2 and LIN28 were not expressed in most patients. Regarding other circulating progenitor cells we observed that PF-PMF had higher mean values of progenitor endothelial cells (PEC) (130±271/ml; range 0-683) than F-PMF (7±16/ml; range 0-37). Mesenchymal progenitor cells (MPC) had also higher mean values in PF-PMF (289±452/ml; range 0-1165) than in F-PMF (111±199/ml; range 0-458). Conversely, Lin-/CD45-CD34+AC133+CXCR4+ cell subset detection was higher in F-PMF (185±381/ml; range 0-882) than in PF-PMF (38±60/ml; range 0-140). Patients V617F-JAK2pos presented lower levels of PEC and MPC (16±19/ml and 43±61/ml respectively) than those V617F-JAK2neg (173±340/ml and 515±470/ml respectively). Similar numbers of Lin-/CD45-CD34+AC133+CXCR4+ cells were observed in both V617F-JAK2pos and V617F-JAK2neg patients (133±331/ml and 123±115/ml respectively).Patients with elevated IPSS (>3) showed higher levels of PEC, MPC and Lin-/CD45-CD34+AC133+CXCR4+ cells (respectively 180±335/ml, 334±560/ml and 256±422/ml) than those with low IPSS (<2) (respectively 12±16/ml, 134±183/ml and 46±92/ml). Based on these findings we were able to define some putative patterns of bloodstream progenitor cell traffic. Thus, PEC appeared to circulate more in PF-PMF, V617F-JAK2neg with a high IPSS. The same pattern was observed regarding MPC. Conversely, Lin-/CD45-CD34+AC133+CXCR4+ cells appeared to circulate more in F-PMF, V617F-JAK2pos with a high IPSS (Table 1). No mutations were detected in the hot-spot of SRSF2 (codons 93-95) which have been associated to a fibrotic progression of PMF.We also performed a bio-informatics analysis of microarray databases comparing the molecular libraries of the gene expression ommibus dataseries GSE3410 and GSE31869 (VSELs (Lin-CD45-CXCR4+CD34-) and CD34+ VSELs (Lin-CD45-CXCR4+CD34+) with the molecular profiling of circulating CD34+ cells from PMF patients and bone marrow CD34+ cells from healthy donors and we observed a tight clustering between PMF and VSELs cells samples (Figure 1).Conclusions. Variations in the blood traffic of some progenitor cell subsets in patients with PMF were observed, including Lin-/CD45-CD34+AC133+CXCR4+ cells. The fact this cell subset was detected in higher number in F-PMF and in patients with a high IPSS, as well as a higher expression of OCT4 in F-PMF supports in part our hypothesis that PMF evolution can be associated to the recruitment and circulation of some primitive progenitors (dormant in the adult life) as it can be observed during the fetal period. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Evidence for Vasculogenic Potential and Endothelial Differentiation of Bone-Marrow-Derived Very Small Embryonic-like Stem Cells

Objective: Very small embryonic-like stem cells (VSELs) are multipotent stem cells localized in adult bone marrow (BM) that may be mobilized into peripheral blood (PB) in response to tissue injury. We aimed to quantify VSELs in BM and PB of patients with critical limb ischemia (CLI) and to test their angiogenic potential in vitro as well as their therapeutic capacity in mouse model of CLI.Approach and Results: We isolated BM VSELs from patients with CLI and studied their potential to differentiate into vascular lineages. Flow and imaging cytometry showed that VSEL counts were lower in BM (p<0.001) and higher (p<0.001) in PB from CLI patients compared to healthy controls, suggesting that ischemia may trigger VSELs mobilization in this patient population. Sorted BM-VSELs cultured in angiogenic media acquired a mesenchymal phenotype (CD90+, Thy-1 gene positive expression). VSEL-derived cells had a pattern of secretion similar to that of endothelial progenitor cells, as they released low levels of VEGF-A and inflammatory cytokines. Noteworthy, VSELs triggered post-ischemic revascularization in immunodeficient mice (p<0.05 vs PBS treatment), and acquired an endothelial phenotype either in vitro when cultured in the presence of VEGF-B (Cdh-5 gene positive expression), or in vivo in Matrigel implants (human CD31+ staining in neo-vessels from plug sections).Conclusions: VSELs are a potential new source of therapeutic cells that may give rise to cells of the endothelial lineage in humans. DisclosuresNo relevant conflicts of interest to declare.

Plasticity of male germline stem cells and their applications in reproductive and regenerative medicine.

Spermatogonial stem cells (SSCs), also known as male germline stem cells, are a small subpopulation of type A spermatogonia with the potential of self-renewal to maintain stem cell pool and differentiation into spermatids in mammalian testis. SSCs are previously regarded as the unipotent stem cells since they can only give rise to sperm within the seminiferous tubules. However, this concept has recently been challenged because numerous studies have demonstrated that SSCs cultured with growth factors can acquire pluripotency to become embryonic stem-like cells. The in vivo and in vitro studies from peers and us have clearly revealed that SSCs can directly transdifferentiate into morphologic, phenotypic, and functional cells of other lineages. Direct conversion to the cells of other tissues has important significance for regenerative medicine. SSCs from azoospermia patients could be induced to differentiate into spermatids with fertilization and developmental potentials. As such, SSCs could have significant applications in both reproductive and regenerative medicine due to their unique and great potentials. In this review, we address the important plasticity of SSCs, with focuses on their self-renewal, differentiation, dedifferentiation, transdifferentiation, and translational medicine studies.

Treatment with embryonic stem-like cells into osteochondral defects in sheep femoral condyles.

BackgroundArticular cartilage has poor intrinsic capacity for regeneration because of its avascularity and very slow cellular turnover. Defects deriving from trauma or joint disease tend to be repaired with fibrocartilage rather than hyaline cartilage. Consequent degenerative processes are related to the width and depth of the defect. Since mesenchymal stem cells (MSCs) deriving from patients affected by osteoarthritis have a lower proliferative and chondrogenic activity, the systemic or local delivery of heterologous cells may enhance regeneration or inhibit the progressive loss of joint tissue. Embryonic stem cells (ESCs) are very promising, since they can self-renew for prolonged periods without differentiation and can differentiate into tissues from all the 3 germ layers. To date only a few experiments have used ESCs for the study of the cartilage regeneration in animal models and most of them used laboratory animals. Sheep, due to their anatomical, physiological and immunological similarity to humans, represent a valid model for translational studies. This experiment aimed to evaluate if the local delivery of male sheep embryonic stem-like (ES-like) cells into osteochondral defects in the femoral condyles of adult sheep can enhance the regeneration of articular cartilage. Twenty-two ewes were divided into 5 groups (1, 2, 6, 12 and 24 months after surgery). Newly formed tissue was evaluated by macroscopic, histological, immunohistochemical (collagen type II) and fluorescent in situ hybridization (FISH) assays.ResultsRegenerated tissue was ultimately evaluated on 17 sheep. Samples engrafted with ES-like cells had significantly better histologic evidence of regeneration with respect to empty defects, used as controls, at all time periods.ConclusionsHistological assessments demonstrated that the local delivery of ES-like cells into osteochondral defects in sheep femoral condyles enhances the regeneration of the articular hyaline cartilage, without signs of immune rejection or teratoma for 24 months after engraftment.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-014-0301-9) contains supplementary material, which is available to authorized users.

The molecular nature of very small embryonic-like stem cells in adult tissues.

Pluripotent stem cells (PSCs) have been considered as the most important cells in regenerative medicine as they are able to differentiate into all types of cells in the human body. PSCs have been established from several sources of embryo tissue or by reprogramming of terminally differentiated adult tissue by transduction of so-called Yamanaka factors (Oct4, Sox2, Klf4, and cMyc). Interestingly, accumulating evidence has demonstrated the residence of PSCs in adult tissue and with the ability to differentiate into multiple types of tissue-committed stem cells (TCSCs). We also recently demonstrated that a population of pluripotent Oct4+ SSEA-1+Sca-1+Lin−CD45− very small embryonic-like stem cells (VSELs) resides in the adult murine bone marrow (BM) and in other murine tissue. These very small (∼3–6 μm) cells express pluripotent markers such as Oct4, Nanog, and SSEA-1. VSELs could be specified into several tissue-residing TCSCs in response to tissue/organ injury, and thus suggesting that these cells have a physiological role in the rejuvenation of a pool of TCSCs under steady-state conditions. In this review article, we discuss the molecular nature of the rare population of VSELs which have a crucial role in regulating the pluripotency, proliferation, differentiation, and aging of these cells.

Isolation and characterization of stem cells in the adult mammalian ovary.

Female mammals are born with a fixed pool of germ cells, which does not replenish during adult life. However, this has been recently challenged and adult ovaries produce oocytes throughout adult life just like sperm in the testes. Evidence is accumulating on the presence of ovarian stem cells, but the need for robust protocols to isolate, identify, further characterize, and subject them to various functionality tests is essential. Knowledge about the function and potential of ovarian stem cells is well demonstrated by various groups, but their true identity remains elusive because of the variability in the approaches used to identify them by different groups. In order to address this we have made attempts to compile our protocols to isolate, identify, characterize, and culture the stem cells using different animal models including human. Two distinct populations of stem cells exist in adult mammalian ovary, including very small embryonic-like stem cells (VSELs) and the progenitors termed ovarian germ stem cells (OGSCs). VSELs are relatively quiescent and undergo asymmetric cell division to give rise to OGSCs, which divide rapidly, occasionally form germ cell nests and undergo meiosis and differentiation into oocytes, which are surrounded by granulosa cells to assemble as primordial follicles.

Primo Vascular System in Human Umbilical Cord and Placenta

The primo vascular system (PVS) has been observed in various animals such as mice, rats, rabbits, dogs, swine, and cow, but not in humans. In this work, we report on the observation of a human PVS on both the epithelial fascia and inside the blood vessels of the umbilical cord (UC). The main morphological characteristics of the primo vessels (PVs) and primo nodes (PNs) from the human UC were in agreement with those of the PVS in various animal organs, including the thicknesses and the transparency of the PVs, the sizes of the PNs, the broken-line arrangement of the rod-shaped nuclei, the sparse distribution of nuclei, and the presence of hollow lumens in the central inner parts of the PNs. It was rather surprising that the human PV was not thicker than the PVs from small animals. The difference between the PVS and blood/lymph vessels was confirmed using immunofluorescence staining of von Willebrand factor, CD31, LYVE-1, and D2-40. The positive expression of the PVS to proliferating cell nuclear antigen, a cell-proliferation marker, was consistent with the recent finding of very small embryonic-like stem cells in the PVS of mice.

FSH-FSHR3-stem cells in ovary surface epithelium: basis for adult ovarian biology, failure, aging, and cancer.

Despite extensive research, genetic basis of premature ovarian failure (POF) and ovarian cancer still remains elusive. It is indeed paradoxical that scientists searched for mutations in FSH receptor (FSHR) expressed on granulosa cells, whereas more than 90% of cancers arise in ovary surface epithelium (OSE). Two distinct populations of stem cells including very small embryonic-like stem cells (VSELs) and ovarian stem cells (OSCs) exist in OSE, are responsible for neo-oogenesis and primordial follicle assembly in adult life, and are modulated by FSH via its alternatively spliced receptor variant FSHR3 (growth factor type 1 receptor acting via calcium signaling and the ERK/MAPK pathway). Any defect in FSH-FSHR3-stem cell interaction in OSE may affect folliculogenesis and thus result in POF. Ovarian aging is associated with a compromised microenvironment that does not support stem cell differentiation into oocytes and further folliculogenesis. FSH exerts a mitogenic effect on OSE and elevated FSH levels associated with advanced age may provide a continuous trigger for stem cells to proliferate resulting in cancer, thus supporting gonadotropin theory for ovarian cancer. Present review is an attempt to put adult ovarian biology, POF, aging, and cancer in the perspective of FSH-FSHR3-stem cell network that functions in OSE. This hypothesis is further supported by the recent understanding that: i) cancer is a stem cell disease and OSE is the niche for ovarian cancer stem cells; ii) ovarian OCT4-positive stem cells are regulated by FSH; and iii) OCT4 along with LIN28 and BMP4 are highly expressed in ovarian cancers.

Very small embryonic-like stem cells are involved in regeneration of mouse pancreas post-pancreatectomy.

IntroductionDespite numerous research efforts, mechanisms underlying regeneration of pancreas remains controversial. Views are divided whether stem cells are involved during pancreatic regeneration or it involves duplication of pre-existing islets or ductal cells or whether pancreatic islet numbers are fixed by birth or they renew throughout life. Pluripotent embryonic stem (ES) and induced pluripotent stem (iPS) cells have been used by several groups to regenerate diabetic mouse pancreas but the beneficial effects are short-lived. It has been suggested that cells obtained after directed differentiation of ES/iPS cells resemble fetal and not their adult counterparts; thus are functionally different and may be of little use to regenerate adult pancreas. A novel population of pluripotent very small embryonic-like stem cells (VSELs) exists in several adult body tissues in both mice and humans. VSELs have been reported in the mouse pancreas, and nuclear octamer-binding transcription factor 4 (OCT-4) positive, small-sized cells have also been detected in human pancreas. VSELs are mobilized into peripheral blood in streptozotocin treated diabetic mice and also in patients with pancreatic cancer. This study aimed to evaluate whether VSELs are involved during regeneration of adult mouse pancreas after partial pancreatectomy.MethodsMice were subjected to partial pancreatectomy wherein almost 70% of pancreas was surgically removed and residual pancreas was studied on Days 1, 3 and 5 post-surgery.ResultsVSELs were detected in Hematoxylin and Eosin stained smears of pancreatic tissue as spherical, small sized cells with a large nucleus surrounded by a thin rim of cytoplasm and could be sorted as LIN-/CD45-/SCA-1+ cells by flow cytometry. Results reveal that although neutrophils with multi-lobed nuclei are mobilized into the pancreas on day 1 after pancreatectomy, by day 5 VSELs with spherical nuclei, high nucleo-cytoplasmic ratio and nuclear OCT-4 are mobilized into the residual pancreas. VSELs undergo differentiation and give rise to PDX-1 and OCT-4 positive progenitors which possibly regenerate both acinar cells and islets.ConclusionsResults provide direct evidence supporting the presence of VSELs in adult mouse pancreas and their role during regeneration. VSELs are an interesting alternative to ES/iPS cells to regenerate a diabetic pancreas in future.

Adult Stem Cells from the Hyaluronic Acid-Rich Node and Duct System Differentiate into Neuronal Cells and Repair Brain Injury

The existence of a hyaluronic acid-rich node and duct system (HAR-NDS) within the lymphatic and blood vessels was demonstrated previously. The HAR-NDS was enriched with small (3.0-5.0 μm in diameter), adult stem cells with properties similar to those of the very small embryonic-like stem cells (VSELs). Sca-1(+)Lin(-)CD45(-) cells were enriched approximately 100-fold in the intravascular HAR-NDS compared with the bone marrow. We named these adult stem cells "node and duct stem cells (NDSCs)." NDSCs formed colonies on C2C12 feeder layers, were positive for fetal alkaline phosphatase, and could be subcultured on the feeder layers. NDSCs were Oct4(+)Nanog(+)SSEA-1(+)Sox2(+), while VSELs were Oct4(+)Nanog(+)SSEA-1(+)Sox2(-). NDSCs had higher sphere-forming efficiency and proliferative potential than VSELs, and they were found to differentiate into neuronal cells in vitro. Injection of NDSCs into mice partially repaired ischemic brain damage. Thus, we report the discovery of potential adult stem cells that may be involved in tissue regeneration. The intravascular HAR-NDS may serve as a route that delivers these stem cells to their target tissues.

Positive effects of prolonged caloric restriction on the population of very small embryonic-like stem cells - hematopoietic and ovarian implications.

BackgroundLow calorie intake, or calorie restriction (CR) without malnutrition, has been demonstrated in several animal species, including mice, to increase both median and maximum lifespan as well as delay reproductive senescence. Our previous work demonstrated a positive correlation between life span and the number of very small embryonic-like stem cells (VSELs) in long living Laron dwarf mice. These animals have very low levels of circulating insulin-like growth factor 1 (IGF-1) in peripheral blood (PB), maintain higher numbers of hematopoietic stem cells (HSPCs) in bone marrow (BM), and display prolonged fecundity compared with wild type littermates. Since CR lowers the level of IGF-1 in PB, we become interested in the effect of CR on the number of VSELs and HSPCs in BM as well as on the morphology of ovaries and testes.MethodsIn our studies four-week-old female and male mice were subjected to CR by employing an alternate-day ad libitum feeding diet for a period of 9 months.ResultsWe observed that mice on CR had a higher number of BM-residing VSELs than control mice fed ad libitum. These changes correlated with higher numbers of HSPCs in BM, spleen, and peripheral blood (PB) as well as with an increase in the number of primordial and primary follicles in ovaries. At the same time, however, no changes were observed in the testes of mice under CR.ConclusionWe conclude that CR positively affects the pool of VSELs in adult tissues and explains the positive effect of CR on longevity.

Erratum to: Very Small Embryonic-Like Stem Cells from the Murine Bone Marrow Differentiate into Epithelial Cells of the Lung

Erratum to: Very Small Embryonic-Like Stem Cells from the Murine Bone Marrow Differentiate into Epithelial Cells of the Lung

Expression of the erythropoietin receptor by germline-derived cells - further support for a potential developmental link between the germline and hematopoiesis.

BackgroundExpressing several markers of migrating primordial germ cells (PGCs), the rare population of quiescent, bone marrow (BM)-residing very small embryonic-like stem cells (VSELs) can be specified like PGCs into hematopoietic stem/progenitor cells (HSPCs). These two properties of VSELs support the possibility of a developmental origin of HSPCs from migrating PGCs.MethodsTo address a potential link between VSELs and germ line cells we analyzed by RT-PCR and FACS expression of erythropoietin receptor (EpoR) on murine bone marrow- and human umbilical cord blood-derived VSELs, murine and human teratocarcinoma cell lines and human ovarian cancer cells. A proper gating strategy and immunostaining excluded from FACS analysis potential contamination by erythroblasts. Furthermore, the transwell chemotaxis assays as well as adhesion and signaling studies were performed to demonstrate functionality of erythropoietin - EpoR axes on these cells.ResultsWe report here that murine and human VSELs as well as murine and human teratocarcinoma cell lines and ovarian cancer cell lines share a functional EpoR.ConclusionsOur data provide more evidence of a potential developmental link between germline cells, VSELs, and HSCs and sheds more light on the developmental hierarchy of the stem cell compartment in adult tissues.

Retraction of: Quiescent very small embryonic-like stem cells resist oncotherapy and can restore spermatogenesis in germ cell depleted mammalian testis.

Stem Cells and DevelopmentVol. 23, No. 12 RetractionFree AccessRetraction of: Quiescent Very Small Embryonic-Like Stem Cells Resist Oncotherapy and Can Restore Spermatogenesis in Germ Cell Depleted Mammalian TestisSandhya Anand, Deepa Bhartiya, Kalpana Sriraman, Hiren Patel, Dhananjay Manjramkar, Ganesh Bakshi, Vandana Dhamankar, and Purna KurkureSandhya AnandSearch for more papers by this author, Deepa BhartiyaSearch for more papers by this author, Kalpana SriramanSearch for more papers by this author, Hiren PatelSearch for more papers by this author, Dhananjay ManjramkarSearch for more papers by this author, Ganesh BakshiSearch for more papers by this author, Vandana DhamankarSearch for more papers by this author, and Purna KurkureSearch for more papers by this authorPublished Online:5 Jun 2014https://doi.org/10.1089/scd.2013.0059.retractAboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail The authors of the article entitled, “Quiescent Very Small Embryonic-Like Stem Cells Resist Oncotherapy and Can Restore Spermatogenesis in Germ Cell Depleted Mammalian Testis (DOI: 10.1089/scd.2013.0059; PMID: 24079711)” which was published in Stem Cells and Development online ahead of print in its unedited format but not yet included in a full issue, have requested a retraction of the article.The study comprised of animal and human studies. Mouse studies showed that pluripotent VSELs in testis are protected from busulphan treatment and can restore spermatogenesis if healthy somatic (sertoli or mesenchymal) cells are transplanted. This study was approved by Animal Ethics Committee at National Institute for Research in Reproductive Health (NIRRH).The human part of the study involved demonstration of similar pluripotent stem cells in testicular biopsy of seven adult azoospermic survivors of various kinds of childhood cancer.This study was approved by Institutional Committee for Stem Cells Research and Therapy (ICSCRT) at NIRRH and by Institute Ethics Committee of Tata Memorial Hospital (TMH). Biopsies were collected after proper informed consent of the study participants following all principles of bioethics and approvals at TMH. However, the authors have discovered they have erred and did not obtain approval from NIRRH Ethics Committee which was mandatory prior to handling the clinical samples.All authors listed on the paper were contacted by Graham Parker, PhD, the Editor-in-Chief of Stem Cells and Development, to ensure their agreement to this retraction. All authors have agreed to the retraction, except Drs. Bakshi and Kurkure who have refused to communicate their agreement with the retraction.Stem Cells and Development is committed to the highest standards of scientific excellence in peer review, and supports the authors' request to retract the article.FiguresReferencesRelatedDetailsCited byActivation of MT1/MT2 to Protect Testes and Leydig Cells against Cisplatin-Induced Oxidative Stress through the SIRT1/Nrf2 Signaling Pathway19 May 2022 | Cells, Vol. 11, No. 10Stem Cells in Regenerative Therapy9 September 2014 Volume 23Issue 12Jun 2014 InformationCopyright 2014, Mary Ann Liebert, Inc.To cite this article:Sandhya Anand, Deepa Bhartiya, Kalpana Sriraman, Hiren Patel, Dhananjay Manjramkar, Ganesh Bakshi, Vandana Dhamankar, and Purna Kurkure.Retraction of: Quiescent Very Small Embryonic-Like Stem Cells Resist Oncotherapy and Can Restore Spermatogenesis in Germ Cell Depleted Mammalian Testis.Stem Cells and Development.Jun 2014.1428-1428.http://doi.org/10.1089/scd.2013.0059.retractPublished in Volume: 23 Issue 12: June 5, 2014PDF download

Karyotype characterization of in vivo- and in vitro-derived porcine parthenogenetic cell lines.

Mammalian haploid cell lines provide useful tools for both genetic studies and transgenic animal production. To derive porcine haploid cells, three sets of experiments were conducted. First, genomes of blastomeres from 8-cell to 16-cell porcine parthenogenetically activated (PA) embryos were examined by chromosome spread analysis. An intact haploid genome was maintained by 48.15% of blastomeres. Based on this result, two major approaches for amplifying the haploid cell population were tested. First, embryonic stem-like (ES-like) cells were cultured from PA blastocyst stage embryos, and second, fetal fibroblasts from implanted day 30 PA fetuses were cultured. A total of six ES-like cell lines were derived from PA blastocysts. No chromosome spread with exactly 19 chromosomes (the normal haploid complement) was found. Four cell lines showed a tendency to develop to polyploidy (more than 38 chromosomes). The karyotypes of the fetal fibroblasts showed different abnormalities. Cells with 19–38 chromosomes were the predominant karyotype (59.48–60.91%). The diploid cells were the second most observed karyotype (16.17%–22.73%). Although a low percentage (3.45–8.33%) of cells with 19 chromosomes were detected in 18.52% of the fetus-derived cell lines, these cells were not authentic haploid cells since they exhibited random losses or gains of some chromosomes. The haploid fibroblasts were not efficiently enriched via flow cytometry sorting. On the contrary, the diploid cells were efficiently enriched. The enriched parthenogenetic diploid cells showed normal karyotypes and expressed paternally imprinted genes at extremely low levels. We concluded that only a limited number of authentic haploid cells could be obtained from porcine cleavage-stage parthenogenetic embryos. Unlike mouse, the karyotype of porcine PA embryo-derived haploid cells is not stable, long-term culture of parthenogenetic embryos, either in vivo or in vitro, resulted in abnormal karyotypes. The porcine PA embryo-derived diploid fibroblasts enriched from sorting might be candidate cells for paternally imprinted gene research.

Reprogramming sertoli cells into pluripotent stem cells.

Induced pluripotent stem cells (iPSCs) have potential applications in the restoration of fertility, regenerative medicine, and animal biotechnology. In this study, we present the induction of iPSCs from mouse Sertoli cells (SCs) by introducing four factors--Oct4, Sox2, Klf4, and c-Myc. As early as day 3 after induction, expression of these factors was detected and typical embryonic stem-like cells began to form. On day 18, these exogenous genes were silenced and colonies were selected according to morphological characteristics. The iPSCs induced from SCs, termed SCiPSCs, strongly expressed pluripotent markers, showed a normal karyotype, and had proliferation and differentiation characteristics similar to those of embryonic stem cells (ESCs), both in vitro and in vivo. Furthermore, exposure of SCiPSCs to nitric oxide (NO) allowed them to maintain pluripotency through the activation of the pluripotent genes Oct4 and Sox2 and upregulation of Nanog expression. Moreover, NO prevented SCiPSCs from undergoing apoptosis by activating the antiapoptotic genes Bcl2 and Bcl2lll, downregulating the proapoptotic genes Bak1 and Casp7, and blocking the activation of the proapoptotic gene Bac. These effects were reversed by exposure to l-NG-monomethylarginine (l-NMMA), a NO inhibitor. These data demonstrate that iPSCs can be generated from SCs and that the self-renewal and pluripotency of SCiPS cells can be maintained in the presence of NO.

New trends and clinical significance of very small embryonic-like stem cells in Ophthalmology

New trends and clinical significance of very small embryonic-like stem cells in Ophthalmology

Very Small Embryonic-Like Stem Cells: A Scientific Debate?

Stem Cells and DevelopmentVol. 23, No. 7 EditorialVery Small Embryonic-Like Stem Cells: A Scientific Debate?Graham C. ParkerGraham C. ParkerSearch for more papers by this authorPublished Online:28 Mar 2014https://doi.org/10.1089/scd.2014.1500AboutSectionsView articleView Full TextPDF/EPUB ToolsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View article"Very Small Embryonic-Like Stem Cells: A Scientific Debate?." Stem Cells and Development, 23(7), pp. 687–688FiguresReferencesRelatedDetailsCited ByCharacterization and Isolation of Very Small Embryonic-like (VSEL) Stem Cells Obtained from Various Human Hematopoietic Cell Sources6 June 2019 | Stem Cell Reviews and Reports, Vol. 15, No. 5Novel Insights into Adult and Cancer Stem Cell Biology Deepa Bhartiya, Hiren Patel, Ranita Ganguly, Ambreen Shaikh, Yashvi Shukla, Diksha Sharma, and Pushpa Singh7 November 2018 | Stem Cells and Development, Vol. 27, No. 22Mouse Bone Marrow VSELs Exhibit Differentiation into Three Embryonic Germ Lineages and Germ & Hematopoietic Cells in Culture9 January 2017 | Stem Cell Reviews and Reports, Vol. 13, No. 2Reply: An update on endometrial stem cells and progenitors by Deepa Bhartiya24 March 2016 | Human Reproduction Update, Vol. 22, No. 4 Volume 23Issue 7Apr 2014 InformationCopyright 2014, Mary Ann Liebert, Inc.To cite this article:Graham C. Parker.Very Small Embryonic-Like Stem Cells: A Scientific Debate?.Stem Cells and Development.Apr 2014.687-688.http://doi.org/10.1089/scd.2014.1500Published in Volume: 23 Issue 7: March 28, 2014PDF download

Comment on: The Proper Criteria for Identification and Sorting of Very Small Embryonic-Like Stem Cells, and Some Nomenclature Issues

Stem Cells and DevelopmentVol. 23, No. 7 Letter to the EditorComment on: The Proper Criteria for Identification and Sorting of Very Small Embryonic-Like Stem Cells, and Some Nomenclature IssuesKrzysztof Szade, Karolina Bukowska-Strakova, Witold Norbert Nowak, Alicja Jozkowicz, and Jozef DulakKrzysztof SzadeSearch for more papers by this author, Karolina Bukowska-StrakovaSearch for more papers by this author, Witold Norbert NowakSearch for more papers by this author, Alicja JozkowiczSearch for more papers by this author, and Jozef DulakSearch for more papers by this authorPublished Online:4 Mar 2014https://doi.org/10.1089/scd.2014.0028AboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View articleFiguresReferencesRelatedDetailsCited byPluripotent Stem Cells in Adult Tissues: Struggling To Be Acknowledged Over Two Decades16 August 2017 | Stem Cell Reviews and Reports, Vol. 13, No. 6Identification of a New Cell Population Constitutively Circulating in Healthy Conditions and Endowed with a Homing Ability Toward Injured Sites12 November 2015 | Scientific Reports, Vol. 5, No. 1Concise Review: Are Stimulated Somatic Cells Truly Reprogrammed into an ES/iPS-Like Pluripotent State? Better Understanding by Ischemia-Induced Multipotent Stem Cells in a Mouse Model of Cerebral InfarctionStem Cells International, Vol. 2015Very Small Embryonic-Like Stem Cells: A Scientific Debate? Graham C. Parker28 March 2014 | Stem Cells and Development, Vol. 23, No. 7 Volume 23Issue 7Apr 2014 InformationCopyright 2014, Mary Ann Liebert, Inc.To cite this article:Krzysztof Szade, Karolina Bukowska-Strakova, Witold Norbert Nowak, Alicja Jozkowicz, and Jozef Dulak.Comment on: The Proper Criteria for Identification and Sorting of Very Small Embryonic-Like Stem Cells, and Some Nomenclature Issues.Stem Cells and Development.Apr 2014.714-716.http://doi.org/10.1089/scd.2014.0028Online Ahead of Editing: March 4, 2014Published in Volume: 23 Issue 7: March 4, 2014PDF download

Conversion of female germline stem cells from neonatal and prepubertal mice into pluripotent stem cells.

Pluripotent stem cells derived from neonatal or adult testes are a useful tool to examine the mechanisms of pluripotency and a resource for cell-based therapies. However, therapies using these cells will only benefit males but not females. Recently, female germline stem cells (FGSCs) were discovered in ovaries. Whether FGSCs can be converted into pluripotent stem cells, similar to spermatogonial stem cells, is unknown. Here, we demonstrate that female embryonic stem-like cells (fESLCs) can be generated within 1 month from the stably proliferating FGSCs cultured in embryonic stem cell (ESC) medium. fESLCs exhibit properties similar to those of ESCs in terms of marker expression and differentiation potential. Thus, our findings suggest that generation of patient-specific fESLCs is feasible and provides a foundation for personalized regenerative applications.