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Oncology Reports Feb 2018The pituitary sex hormones (SexHs): follicle‑stimulating hormone (FSH), luteinizing hormone (LH), and prolactin (PRL) regulate several functions crucial for...
The pituitary sex hormones (SexHs): follicle‑stimulating hormone (FSH), luteinizing hormone (LH), and prolactin (PRL) regulate several functions crucial for reproduction, including oogenesis, spermatogenesis, and lactation. An important source of prolactin-like hormones, known as lactogens, is the placenta, and lactogens bind to the PRL receptor (PRLR) with high affinity and thereby mimic the actions of PRL. Recently, it has been demonstrated that pituitary SexHs were involved in metastatic lung cancer, certain sarcomas, and leukemia. In the present study we aimed to investigate whether FSH, LH, and PRL were able to stimulate stem cells involved in early development. To address this issue we employed a murine embryonic stem cell line (ES-D3) as well as two teratocarcinoma cell lines, P19 (murine) and NTera2 (human). We determined that all these cells expressed SexH receptors at the mRNA and protein levels and that stimulation of these receptors induced phosphorylation of p42/44 MAPK, p38 MAPK, and AKT. Moreover, ES-D3, P19, and NTera2 cells responded with increased migration and adhesion to physiological concentrations of pituitary SexHs. In view of these findings we proposed that maternal-derived pituitary SexHs regulate the biology of stem cells involved in early development.
Topics: Animals; Cell Adhesion; Cell Movement; Cell Proliferation; Embryonic Stem Cells; Follicle Stimulating Hormone; Gonadotropins, Pituitary; Humans; Luteinizing Hormone; Male; Mice; Prolactin; Receptors, Gonadotropin; Signal Transduction; Teratocarcinoma; Testicular Neoplasms
PubMed: 29207191
DOI: 10.3892/or.2017.6108 -
International Journal of Molecular... Nov 2021About 8% of our genome is composed of sequences from Human Endogenous Retroviruses (HERVs). The HERV-K (HML.2) family, here abbreviated HML.2, is able to produce virus...
About 8% of our genome is composed of sequences from Human Endogenous Retroviruses (HERVs). The HERV-K (HML.2) family, here abbreviated HML.2, is able to produce virus particles that were detected in cell lines, malignant tumors and in autoimmune diseases. Parameters and properties of HML.2 released from teratocarcinoma cell lines GH and Tera-1 were investigated in detail. In most experiments, analyzed viruses were purified by density gradient centrifugation. HML.2 structural proteins, reverse transcriptase (RT) activity, viral RNA (vRNA) and particle morphology were analyzed. The HML.2 markers were predominantly detected in fractions with a buoyant density of 1.16 g/cm. Deglycosylation of TM revealed truncated forms of transmembrane (TM) protein. Free virions and extracellular vesicles (presumably microvesicles-MVs) with HML.2 elements, including budding intermediates, were detected by electron microscopy. Viral elements and assembled virions captured and exported by MVs can boost specific immune responses and trigger immunomodulation in recipient cells. Sequencing of cDNA clones demonstrated exclusive presence of HERV-K108 in HML.2 from Tera-1 cells. Not counting two recombinant variants, four known sequences were found in HML.2 from GH cells. Obtained results shed light on parameters and morphology of HML.2. A possible mechanism of HML.2-induced diseases is discussed.
Topics: Capsid; Cell Line, Tumor; Cell Membrane; Centrifugation, Density Gradient; Endogenous Retroviruses; Extracellular Vesicles; Gene Products, env; Genes, env; HEK293 Cells; Humans; Membrane Proteins; RNA, Viral; Reverse Transcriptase Polymerase Chain Reaction; Teratocarcinoma; Transfection; Viral Envelope; Virus Assembly
PubMed: 34830279
DOI: 10.3390/ijms222212398 -
Stem Cells Translational Medicine May 2016Human induced pluripotent stem cells (iPSCs) and derived progeny provide invaluable regenerative platforms, yet their clinical translation has been compromised by their...
UNLABELLED
Human induced pluripotent stem cells (iPSCs) and derived progeny provide invaluable regenerative platforms, yet their clinical translation has been compromised by their biosafety concern. Here, we assessed the safety of transplanting patient-derived iPSC-generated pancreatic endoderm/progenitor cells. Transplantation of progenitors from iPSCs reprogrammed by lentiviral vectors (LV-iPSCs) led to the formation of invasive teratocarcinoma-like tumors in more than 90% of immunodeficient mice. Moreover, removal of primary tumors from LV-iPSC progeny-transplanted hosts generated secondary and metastatic tumors. Combined transgene-free (TGF) reprogramming and elimination of residual pluripotent cells by enzymatic dissociation ensured tumor-free transplantation, ultimately enabling regeneration of type 1 diabetes-specific human islet structures in vivo. The incidence of tumor formation in TGF-iPSCs was titratable, depending on the oncogenic load, with reintegration of the cMYC expressing vector abolishing tumor-free transplantation. Thus, transgene-free cMYC-independent reprogramming and elimination of residual pluripotent cells are mandatory steps in achieving transplantation of iPSC progeny for customized and safe islet regeneration in vivo.
SIGNIFICANCE
Pluripotent stem cell therapy for diabetes relies on the safety as well as the quality of derived insulin-producing cells. Data from this study highlight prominent tumorigenic risks of induced pluripotent stem cell (iPSC) products, especially when reprogrammed with integrating vectors. Two major underlying mechanisms in iPSC tumorigenicity are residual pluripotent cells and cMYC overload by vector integration. This study also demonstrated that combined transgene-free reprogramming and enzymatic dissociation allows teratoma-free transplantation of iPSC progeny in the mouse model in testing the tumorigenicity of iPSC products. Further safety assessment and improvement in iPSC specification into a mature β cell phenotype would lead to safe islet replacement therapy for diabetes.
Topics: Adult; Aged; Animals; Cell Differentiation; Cells, Cultured; Cellular Reprogramming Techniques; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type 2; Gene Expression Regulation, Neoplastic; Genetic Vectors; Heterografts; Humans; Induced Pluripotent Stem Cells; Insulin; Islets of Langerhans; Islets of Langerhans Transplantation; Keratinocytes; Lentivirus; Male; Mice, SCID; Phenotype; Proto-Oncogene Proteins c-myc; Regeneration; Teratocarcinoma; Transfection
PubMed: 26987352
DOI: 10.5966/sctm.2015-0017 -
International Journal of Molecular... Feb 2021Cripto-1 is a member of the EGF-CFC/FRL1/Cryptic family and is involved in embryonic development and carcinogenesis. We designed a novel anti-Cripto-1 artificial...
Cripto-1 is a member of the EGF-CFC/FRL1/Cryptic family and is involved in embryonic development and carcinogenesis. We designed a novel anti-Cripto-1 artificial antibody and assessed the recognition to the antigen and the potential to suppress the growth of cancer stem cells. First, single chain antibody clones were isolated by bio-panning with the affinity to recombinant Cripto-1 protein from our original phage-display library. Then, the variable regions of heavy chain VH and light chain VL in each clone were fused to constant regions of heavy chain CH and light chain CL regions respectively. These fused genes were expressed in ExpiCHO-S cells to produce artificial humanized antibodies against Cripto-1. After evaluation of the expression levels, one clone was selected and the anti-Cripto-1 antibody was produced and purified. The purified antibody showed affinity to recombinant Cripto-1 at 1.1 pmol and immunoreactivity to cancer tissues and cell lines. The antibody was available to detect the immunoreactivity in tissue microarrays of malignant tumors as well as in Cripto-1 overexpressing cells. Simultaneously, the antibody exhibited the potential to suppress the growth of human colon cancer derived GEO cells overexpressing Cripto-1 with IC at approximately 110 nM. The artificially humanized antibody is proposed to be a good candidate to target cancer cells overexpressing Cripto-1.
Topics: Amino Acid Sequence; Antibodies, Monoclonal, Humanized; Antineoplastic Agents; Breast Neoplasms; Cell Proliferation; Female; GPI-Linked Proteins; Humans; Intercellular Signaling Peptides and Proteins; Male; Neoplasm Proteins; Sequence Homology; Teratocarcinoma; Testicular Neoplasms; Tumor Cells, Cultured
PubMed: 33567764
DOI: 10.3390/ijms22041709 -
Microorganisms Oct 2023Natural metabolites from beneficial fungi were recognized for their potential to inhibit multidrug-resistant human and plant fungal pathogens. The present study...
Natural metabolites from beneficial fungi were recognized for their potential to inhibit multidrug-resistant human and plant fungal pathogens. The present study describes the isolation, metabolite profiling, antibacterial, and antifungal, antioxidant, and anticancer activities of soil fungi. Among the 17 isolates, the AK-7 isolate was selected based on the primary screening. Further, the identification of isolate AK-7 was performed by 18S rRNA sequencing and identified as (with 99.90% similarity). Additionally, the ethyl acetate extract of the strain AK-7 (AK-7 extract) was characterized by Fourier Transform Infrared Spectroscopy (FTIR) and a Gas Chromatography-Mass Spectroscopy (GC-MS) analysis, and the results showed different functional groups and bioactive metabolites. Consequently, a secondary screening of antibacterial activity by the agar well diffusion method showed significant antibacterial activity against Gram-negative and Gram-positive bacterial pathogens. The AK-7 extract exhibited notable antifungal activity by a food poisoning method and showed maximum inhibition of 77.84 ± 1.62%, 56.42 ± 1.27%, and 37.96 ± 1.84% against , and phytopathogens. Consequently, the AK-7 extract showed significant antioxidant activity against DPPH and ABTS free radicals with IC values of 59.084 μg/mL and 73.36 μg/mL. Further, the anticancer activity of the AK-7 extract against the human ovarian teratocarcinoma (PA-1) cell line was tested by MTT and Annexin V flow cytometry. The results showed a dose-dependent reduction in cell viability and exhibited apoptosis with an IC value of 82.04 μg/mL. The study highlights the potential of the strain AK-7 as a source of active metabolites and natural antibacterial, antifungal, antioxidant, and anticancer agent, and it could be an excellent alternative for pharmaceutical and agricultural sectors.
PubMed: 37894138
DOI: 10.3390/microorganisms11102480 -
Biochimica Et Biophysica Acta May 2005Embryonic stem (ES) cells are pluripotent, possessing the unique property to differentiate into any somatic cell type while retaining the ability to proliferate... (Review)
Review
Embryonic stem (ES) cells are pluripotent, possessing the unique property to differentiate into any somatic cell type while retaining the ability to proliferate indefinitely. Due to their ability to recapitulate embryonic differentiation, ES cells are an ideal tool to study the process of early embryogenesis in vitro. Signalling cascades and genes involved in differentiation can be easily studied, and functional genomics approaches aim to identify the regulatory networks underlying lineage commitment. Their unique ability to differentiate into any cell type make ES cells a prime candidate for cell replacement therapy (CRT) of various degenerative disorders. Results from various disease models are promising and have demonstrated their principal suitability as a therapeutic agent in diseases such as myocardial infarctions, diabetes mellitus and Parkinson's disease. Prior to clinical trials in humans, two issues remain to be solved: due to their high proliferative potential, ES cells can form teratocarcinomas in the recipient, and depending on the source of the cells, ES cell grafts may be rejected by the host organism. This review discusses the current state of basic ES cell research with a focus on cardiac differentiation and gives an overview of their use in CRT approaches.
Topics: Animals; Cardiology; Cell Differentiation; Cell Proliferation; Embryo Research; Embryo, Mammalian; Heart; Humans; Myocardium; Pluripotent Stem Cells; Research Design; Stem Cell Transplantation; Stem Cells
PubMed: 15949691
DOI: 10.1016/j.bbadis.2004.11.018 -
Frontiers in Chemistry 2022The biogenic approach of synthesizing metal nanoparticles is an exciting and interesting research area with a wide range of applications. The present study reports a...
The biogenic approach of synthesizing metal nanoparticles is an exciting and interesting research area with a wide range of applications. The present study reports a simple, convenient, low-cost method for synthesizing magnesium oxide nanoparticles (MgONPs) from pumpkin seed extracts and their anticancer efficacy against ovarian teratocarcinoma cell line (PA-1). The characteristic features of biogenic MgONPs were assessed by UV-visible spectrophotometry (UV-vis), X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The formation of spherical NPs with an average size of 100 nm was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Moreover, MgONPs exhibit considerable cytotoxicity with an IC dose of 12.5 μg/ml. A dose-dependent rise in the induction of apoptosis, ROS formation, and inhibition in the migration of PA-1 cells was observed up to 15 μg/ml concentration, reflecting their significant anticancer potential against ovarian teratocarcinoma cell line. However, additional work, especially in different and models, is recommended to find out their real potential before this environment-friendly and cost-effective nanoformulation could be exploited for the benefit of humankind.
PubMed: 36186592
DOI: 10.3389/fchem.2022.970193 -
Methods in Molecular Biology (Clifton,... 2013This chapter describes the culture and propagation of murine embryonic stem cells, F9 and P19 and strategies for differentiation of these stem cells into neurons....
This chapter describes the culture and propagation of murine embryonic stem cells, F9 and P19 and strategies for differentiation of these stem cells into neurons. Protocols focus on maintenance and propagation of these cells and routine procedures employed for differentiation into neuronal cells. Additional protocols are also described for obtaining enriched populations of mature neurons from P19 cells and differentiation of F9 cells into serotonergic or catecholaminergic neurons.The protocols described herein can be employed for dissection of the pathways such as gliogenesis and neurogenesis that are involved in differentiation of pluripotent stem cells such as F9 and P19 into glial cells or terminally differentiated neurons.
Topics: Animals; Catecholamines; Cell Culture Techniques; Cell Differentiation; Cell Line, Tumor; Cell Proliferation; Cryopreservation; Mice; Neurons; Serotonergic Neurons; Teratocarcinoma
PubMed: 23975819
DOI: 10.1007/978-1-62703-640-5_4 -
Oncotarget Aug 2016Glutamate behaves as the principal excitatory neurotransmitter in the vertebrate central nervous system and recently demonstrates intercellular signaling activities in...
Glutamate behaves as the principal excitatory neurotransmitter in the vertebrate central nervous system and recently demonstrates intercellular signaling activities in periphery cancer cells. How the glutamatergic transmission is organized and operated in cancer stem cells remains undefined. We have identified a glutamatergic transmission circuit in embryonal carcinoma stem cells. The circuit is organized and operated in an autocrine mechanism and suppresses the cell proliferation and motility. Biological analyses determined a repertoire of glutamatergic transmission components, glutaminase, vesicular glutamate transporter, glutamate NMDA receptor, and cell membrane excitatory amino-acid transporter, for glutamate biosynthesis, package for secretion, reaction, and reuptake in mouse and human embryonal carcinoma stem cells. The glutamatergic components were also identified in mouse transplanted teratocarcinoma and in human primary teratocarcinoma tissues. Released glutamate acting as the signal was directly quantified by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Genetic and pharmacological abolishment of the endogenously released glutamate-induced tonic activation of the NMDA receptors increased the cell proliferation and motility. The finding suggests that embryonal carcinoma stem cells can be actively regulated by establishing a glutamatergic autocrine/paracrine niche via releasing and responding to the transmitter.
Topics: Animals; Autocrine Communication; Cell Line, Tumor; Cell Movement; Cell Proliferation; Chromatography, Liquid; Cytosol; Embryonal Carcinoma Stem Cells; Gene Expression Regulation, Neoplastic; Glutamic Acid; Glutamine; Humans; Mice; Mice, Inbred BALB C; Neurotransmitter Agents; Receptors, N-Methyl-D-Aspartate; Tandem Mass Spectrometry
PubMed: 27322683
DOI: 10.18632/oncotarget.9973 -
The American Journal of Pathology Aug 1981The origin of the endocrine cells in the respiratory tract and the gastrointestinal tract is still a matter of debate. In the original concept of the amine precursor... (Comparative Study)
Comparative Study
The origin of the endocrine cells in the respiratory tract and the gastrointestinal tract is still a matter of debate. In the original concept of the amine precursor uptake and decarboxylation (APUD) system, all APUD cells were considered to be derived from the neural crest. More recently it has been proposed that the APUD cell types of the gastrointestinal and respiratory tracts originate from neuroendocrine-programmed ectoblast. Still other investigators have reported observations that favor a direct endodermal origin of these cell types. Based on the assumption that in teratomas different tissue types which in normal embryogenesis are derived from the neuroectoderm might be expected to occur together, we investigated a series of cystic ovarian teratomas and testicular teratocarcinomas for the presence of brain tissue and of different types of APUD cells. In the ovarian teratomas, intestinal and respiratory APUD cell types were found almost exclusively without coexistence of brain tissue, whereas melanocytes, which are of neuroectodermal origin, occurred mostly together with brain tissue. In the testicular teratocarcinomas, intestinal types of APUD cells occurred without brain tissue. Peptide hormone production was found in appropriate tissues. It can therefore be concluded that in teratomas appropriate intestinal and respiratory APUD cells differentiate in and presumably descend directly from intestinal and respiratory epithelium.
Topics: APUD Cells; Cell Differentiation; Female; Humans; Male; Melanocytes; Ovarian Neoplasms; Teratoma; Testicular Neoplasms
PubMed: 6114639
DOI: No ID Found