Oxygen modulates human embryonic stem cell metabolism in the absence of changes in self-renewal

2016 ◽  
Vol 28 (4) ◽  
pp. 446 ◽  
Author(s):  
Alexandra J. Harvey ◽  
Joy Rathjen ◽  
Lijia Jackie Yu ◽  
David K. Gardner
Keyword(s):  
Stem Cell ◽  
Es Cells ◽  
Atmospheric Oxygen ◽  
Embryonic Stem ◽  
Lactate Production ◽  
Cell Physiology ◽  
Stem Cell Markers ◽  
Es Cell ◽  
Oxygen Conditions ◽  
Cells Cultured

Human embryonic stem (ES) cells are routinely cultured under atmospheric oxygen (~20%), a concentration that is known to impair embryo development in vitro and is likely to be suboptimal for maintaining human ES cells compared with physiological (~5%) oxygen conditions. Conflicting reports exist on the effect of oxygen during human ES cell culture and studies have been largely limited to characterisation of typical stem cell markers or analysis of global expression changes. This study aimed to identify physiological markers that could be used to evaluate the metabolic impact of oxygen on the MEL-2 human ES cell line after adaptation to either 5% or 20% oxygen in extended culture. ES cells cultured under atmospheric oxygen displayed decreased glucose consumption and lactate production when compared with those cultured under 5% oxygen, indicating an overall higher flux of glucose through glycolysis under physiological conditions. Higher glucose utilisation at 5% oxygen was accompanied by significantly increased expression of all glycolytic genes analysed. Analysis of amino acid turnover highlighted differences in the consumption of glutamine and threonine and in the production of proline. The expression of pluripotency and differentiation markers was, however, unaltered by oxygen and no observable difference in proliferation between cells cultured in 5% and 20% oxygen was seen. Apoptosis was elevated under 5% oxygen conditions. Collectively these data suggest that culture conditions, including oxygen concentration, can significantly alter human ES cell physiology with coordinated changes in gene expression, in the absence of detectable alterations in undifferentiated marker expression.

Derivation, Characterization, and In Vitro Differentiation of Canine Embryonic Stem Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4059-4059
Author(s):  
Aravind Ramakrishnan ◽  
Brian Hayes ◽  
Sara R. Fagerlie ◽  
Szczepan Baran ◽  
Michael Harkey ◽  
...  
Keyword(s):  
Stem Cell ◽  
Es Cells ◽  
Embryonic Stem Cell Line ◽  
Embryonic Stem ◽  
Embryoid Bodies ◽  
Clara Cell ◽  
In Vivo Model ◽  
Large Animal ◽  
Es Cell

Abstract Embryonic stem (ES) cells have created considerable excitement in the last few years due to their unlimited potential to produce cells for tissue repair and replacement. However, a large animal pre-clinical model is necessary to establish the safety and efficacy of ES cell-derived tissue replacement therapy. The canine model has long been used in medical research, has been well established to study adult stem cell transplantation and has been highly predictive of clinical outcomes in humans, more so than rodent models. Given the documented record for extrapolating from dog to man, we hypothesize that the dog would serve as an ideal pre-clinical in vivo model for studying the clinical applications of ESC derived tissue. Eleven putative ES cell lines were initiated from canine blastocysts harvested from natural matings. One line described here, FHDO-7, has been maintained through 34 passages and has many characteristics of ES cells from other species. FHDO-7 cells are alkaline phosphatase positive and express both message and protein for the Oct4 transcription factor. They also express message for Nanog and do not express message for Cdx2 which is associated with trophectoderm. Furthermore, they express a cluster of pluripotency-associated microRNAs (miR-302b, miR-302c and miR-367) that have been found to be characteristic of human and mouse ES cells. The FHDO-7 cells grow on feeder layers of modified mouse embryonic fibroblasts (MEF) as flat colonies that resemble ES cells from mink, a close phylogenetic relative of dog. When cultured in nonadherent plates without feeders the cells form embryoid bodies (EB). Under various culture conditions the EBs give rise to ectoderm-derived neuronal cells expressing β3-tubulin, mesoderm-derived osteocytes producing bone, and endoderm-derived cells expressing alpha feto protein or Clara cell specific protein. These results indicate that FHDO-7 is a pluripotent embryonic stem cell line.

Transcriptional profiling of reporter genes used for molecular imaging of embryonic stem cell transplantation

2006 ◽  
Vol 25 (1) ◽  
pp. 29-38 ◽  
Author(s):  
Joseph C. Wu ◽  
Joshua M. Spin ◽  
Feng Cao ◽  
Shuan Lin ◽  
Xiaoyan Xie ◽  
...  
Keyword(s):  
Stem Cell ◽  
Molecular Imaging ◽  
Cell Fate ◽  
Reporter Gene ◽  
Lentiviral Vector ◽  
Es Cells ◽  
Embryonic Stem ◽  
Reporter Genes ◽  
Nucleic Acid Metabolism ◽  
Es Cell

Stem cell therapy offers exciting promise for treatment of ischemic heart disease. Recent advances in molecular imaging techniques now allow investigators to monitor cell fate noninvasively and repetitively. Here we examine the effects of a triple-fusion reporter gene on embryonic stem (ES) cell transcriptional profiles. Murine ES cells were stably transfected with a self-inactivating lentiviral vector carrying a triple-fusion (TF) construct consisting of fluorescence, bioluminescence, and positron emission tomography (PET) reporter genes. Fluorescence-activated cell sorting (FACS) analysis allowed isolation of stably transfected populations. Microarray studies comparing gene expression in nontransduced control ES cells vs. stably transduced ES cells expressing triple fusion (ES-TF) revealed some increases in transcriptional variability. Annotation analysis showed that ES-TF cells downregulated cell cycling, cell death, and protein and nucleic acid metabolism genes while upregulating homeostatic and anti-apoptosis genes. Despite these transcriptional changes, expression of the TF reporter gene had no significant effects on ES cell viability, proliferation, and differentiation capability. Importantly, transplantation studies in murine myocardium demonstrated the feasibility of tracking ES-TF cells in living subjects using bioluminescence and PET imaging. Taken together, this is the first study to analyze in detail the effects of reporter genes on molecular imaging of ES cells.

A Study on the Induction of Mesoderm Differentiation of Mouse Embryonic Stem Cell in Serum Free Culture.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4161-4161
Author(s):  
Takao Suzuki ◽  
Masatomo Takahashi ◽  
Tsukasa Miyagi ◽  
Michiko Irei ◽  
Yasushi Shibuya ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Es Cells ◽  
Calf Serum ◽  
Embryonic Stem ◽  
Feeder Cells ◽  
Es Cell ◽  
Rt Pcr ◽  
Serum Free

Abstract (Introduction) Several products which can maintain the growth of ES (Embryonic stem) cell with no feeder cells or fetal calf serum (FCS) is now commercially available. Culti Cell ®(CC) can be a substitute for FCS and has ability of mainitaining mouse ES cells without feeder cells. Previously, it was reported that ES cell differentiate into FLk (fetal liver kinase) positive cells derived from mesoderm in methylcellulose and FCS. We investigated whether ES cell is able to differentiate into Flk positive cell in CC and methylcellulose without FCS. (Materials and Methods) Thawed ES cells were cultured with methylcellulose and CC in the absence of FCS and feeder cells. EB (Embryoid body) formation was completed after 7 days. The separated cell from EB was adjusted to 1×104/ml. These ES cells were cultured with 15% CC and 0.8% methylcellulose for 7 days at 37°C in 5% CO2. The expression of Flk and mesoderm differentiation was analyzed with flow cytometry and reverse transcriptase-polymerase chain reaction (RT-PCR) at day 1, 2, 3, 5 and 7. Oligonucleotide primers for Flk1 and brachyury were used for RT-PCR. In addition, the expression of CD31 was also examined with flow cytometry. (Results) No Flk positive cells were detected in these culture periods. The expression of brachyury, which is a marker of mesoderm differentiation, was consistently observed throughout culture days. The expression of CD31 gradually diminished as the culture period increased. (Conclusion) These results suggest that ES cell is maintained and differentiate into mesoderm in suspension culture with CC and methylcellulose, but diferentiation to Flk positive cell can not be induced in this culture condition. Further examination is necessary to clarify unidentified factors for differentiation to Flk positive cell in serum free culture.

Maintenance of pluripotential embryonic stem cells by stem cell selection

1998 ◽  
Vol 10 (8) ◽  
pp. 527 ◽  
Author(s):  
Peter Mountford ◽  
Jennifer Nichols ◽  
Branko Zevnik ◽  
Carmel O'Brien ◽  
Austin Smith
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Es Cells ◽  
Embryonic Stem ◽  
Embryoid Bodies ◽  
Stem Cell Population ◽  
Cell Selection ◽  
Es Cell ◽  
Mammalian Embryo ◽  
Differentiated Cells

As gastrulation proceeds, pluripotential stem cells with the capacity to contribute to all primary germ layers disappear from the mammalian embryo. The extinction of pluripotency also occurs during the formation of embryoid bodies from embryonic stem (ES) cells. In this report we show that if the initial differentiated progeny are removed from ES cell aggregates, further differentiation does not proceed and the stem cell population persists and expands. Significantly, the presence of even minor populations of differentiated cells lead to the complete loss of stem cells from the cultures. This finding implies that the normal elimination of pluripotent cells is dictated by inductive signals provided by differentiated progeny. We have exploited this observation to develop a strategy for the isolation of pluripotential cells. This approach, termed stem cell selection, may have widespread applicability to the derivation and propagation of stem cells.

scholarly journals Embryonic Stem Cell Lines of Nonhuman Primates

2002 ◽  
Vol 2 ◽  
pp. 1762-1773 ◽  
Author(s):  
Norio Nakatsuji ◽  
Hirofumi Suemori
Keyword(s):  
Stem Cell ◽  
Cell Therapy ◽  
Cell Lines ◽  
Nonhuman Primate ◽  
Cynomolgus Monkey ◽  
Es Cells ◽  
Embryonic Stem ◽  
Embryoid Bodies ◽  
Primate Species ◽  
Es Cell

Human embryonic stem (ES) cell lines have opened great potential and expectation for cell therapy and regenerative medicine. Monkey and human ES cell lines, which are very similar to each other, have been established from monkey blastocysts and surplus human blastocysts from fertility clinics.Nonhuman primate ES cell lines provide important research tools for basic and applicative research. Firstly, they provide wider aspects of investigation of the regulative mechanisms of stem cells and cell differentiation among primate species. Secondly, their usage does not need clearance or permission from the regulative rules in many countries that are associated with the ethical aspects of human ES cells, although human and nonhuman embryos and fetuses are very similar to each other. Lastly and most importantly, they are indispensable for animal models of cell therapy to test effectiveness, safety, and immunological reaction of the allogenic transplantation in a setting similar to the treatment of human diseases.So far, ES cell lines have been established from rhesus monkey (Macaca mulatta), common marmoset (Callithrix jacchus), and cynomolgus monkey (Macaca fascicularis), using blastocysts produced naturally or by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). These cell lines seem to have very similar characteristics. They express alkaline phosphatase activity and stage-specific embryonic antigen (SSEA)-4 and, in most cases, SSEA-3. Their pluripotency was confirmed by the formation of embryoid bodies and differentiation into various cell types in culture and also by the formation of teratomas that contained many types of differentiated tissues including derivatives of three germ layers after transplantation into the severe combined immunodeficiency (SCID) mice.The noneffectiveness of the leukemia inhibitory factor (LIF) signal makes culture of primate and human ES cell lines prone to undergo spontaneous differentiation and thus it is difficult to maintain these stem cell colonies. Also, these ES cells are more susceptible to various stresses, causing difficulty with subculturing using enzymatic treatment and cloning from single cells. However, with various improvements in culture methods, it is now possible to maintain stable colonies of monkey ES cells using a serum-free medium and subculturing with trypsin treatment. Under such conditions, cynomolgus monkey ES cell lines can be maintained in an undifferentiated state with a normal karyotype and pluripotency even after prolonged periods of culture over 1 year. Such progress should facilitate many aspects of stem cell research using both nonhuman primate and human ES cell lines.

EU patent changes may spur stem cell research

2015 ◽  
Keyword(s):  
Stem Cell ◽  
Stem Cell Research ◽  
Es Cells ◽  
Public Support ◽  
Embryonic Stem ◽  
Commercial Application ◽  
Es Cell ◽  
Content Type ◽  
Eu Member States ◽  
European Court

Subject The implications of the ECJ ruling on embryonic stem cell research. Significance The European Court of Justice (ECJ) on December 18 reversed part of its general ban on patents for human embryonic stem (ES) cells. ES cells are capable of developing into any type of cell. Their potential therapeutic application is limitless and human ES cells could be used to treat diseases ranging from diabetes and stroke, to Parkinson's and Alzheimer's disease. Patenting not only the technology for producing human ES cell-based therapies, but the cells themselves, broadens the scope of protection and commercial application -- enabling EU member states to compete with the rest of the world. Impacts The 2014 ruling opens the door for further challenges to the general ban on human ES-cell patents of 2011. The latest ruling somewhat undercuts opposition to human ES cell research on ethical grounds. Removing the stigma of immorality may increase public support and funding for human ES cell research.

Chromatin regulation landscape of embryonic stem cell identity

2010 ◽  
Vol 31 (2) ◽  
pp. 77-86 ◽  
Author(s):  
Yun Hwa Lee ◽  
Qiang Wu
Keyword(s):  
Stem Cell ◽  
Inner Cell Mass ◽  
Es Cells ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Histone Variants ◽  
Transcriptional Networks ◽  
Chromatin Regulation ◽  
Es Cell ◽  
Cell Identity

ES cells (embryonic stem cells) derived from the ICM (inner cell mass) of blastocysts are pluripotent and are capable of giving rise to most cell types. The ES cell identity is mainly maintained by the Oct4 (octamer-binding transcription factor 4) and Nanog transcriptional networks. Recently, a tremendous amount of work has focused on deciphering how ES cell identity is regulated epigenetically. It has been shown that histone methylation/demethylation, histone acetylation/deacetylation, histone variants and chromatin remodelling play crucial roles in ES cell maintenance and differentiation. Moreover, perturbation of those chromatin regulators results in loss of ES cell identity or aberrant differentiation. Therefore, it is important to fully understand the chromatin regulation landscape of ES cells. The knowledge gained will help us to harness the unique characteristics of ES cells for stem cell-related therapy and regenerative medicine. In the present review, we will discuss recent proceedings that provide novel insights into chromatin regulation of ES cell identity.

The expression of mesenchymal and embryonic stem cell markers by human limbal Stromal cells

2011 ◽  
Author(s):  
Moon Nian Lim ◽  
Umapathy Thiageswari ◽  
Othman Ainoon ◽  
P. J. N. Baharuddin ◽  
R. A. Jamal ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Stromal Cells ◽  
Embryonic Stem ◽  
Stem Cell Markers ◽  
Cell Markers ◽  
Embryonic Stem Cell Markers

scholarly journals PRL3 induces polypoid giant cancer cells eliminated by PRL3-zumab to reduce tumor relapse

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Min Thura ◽  
Zu Ye ◽  
Abdul Qader Al-Aidaroos ◽  
Qiancheng Xiong ◽  
Jun Yi Ong ◽  
...  
Keyword(s):  
Dna Damage ◽  
Stem Cell ◽  
Cancer Cells ◽  
Embryonic Stem ◽  
Genotoxic Stress ◽  
Stem Cell Markers ◽  
Tumor Removal ◽  
Primary Tumors ◽  
Tumor Relapse ◽  
Dna Damage Signaling

AbstractPRL3, a unique oncotarget, is specifically overexpressed in 80.6% of cancers. In 2003, we reported that PRL3 promotes cell migration, invasion, and metastasis. Herein, firstly, we show that PRL3 induces Polyploid Giant Cancer Cells (PGCCs) formation. PGCCs constitute stem cell-like pools to facilitate cell survival, chemo-resistance, and tumor relapse. The correlations between PRL3 overexpression and PGCCs attributes raised possibilities that PRL3 could be involved in PGCCs formation. Secondly, we show that PRL3+ PGCCs co-express the embryonic stem cell markers SOX2 and OCT4 and arise mainly due to incomplete cytokinesis despite extensive DNA damage. Thirdly, we reveal that PRL3+ PGCCs tolerate prolonged chemotherapy-induced genotoxic stress via suppression of the pro-apoptotic ATM DNA damage-signaling pathway. Fourthly, we demonstrated PRL3-zumab, a First-in-Class humanized antibody drug against PRL3 oncotarget, could reduce tumor relapse in ‘tumor removal’ animal model. Finally, we confirmed that PGCCs were enriched in relapse tumors versus primary tumors. PRL3-zumab has been approved for Phase 2 clinical trials in Singapore, US, and China to block all solid tumors. This study further showed PRL3-zumab could potentially serve an ‘Adjuvant Immunotherapy’ after tumor removal surgery to eliminate PRL3+ PGCC stem-like cells, preventing metastasis and relapse.

Sign in / Sign up

Export Citation Format

Share Document