scholarly journals 172 PRIMORDIAL GERM CELL DIFFERENTIATION FROM ES CELLS IN VITRO IN MOUSE

2005 ◽  
Vol 17 (2) ◽  
pp. 236
Author(s):  
J. Kobolak ◽  
E. Deak ◽  
A. Dinnyes
Keyword(s):  
Gene Expression ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Expression Profiles ◽  
Es Cells ◽  
Embryoid Bodies ◽  
Es Cell ◽  
Sodium Pyruvate ◽  
Germ Cell Differentiation

The primordial germ cells (PGC) in the genital ridge of the embryo are the progenitors of sperm and eggs. The goal of the present study was to derive PGC cells from embryonic stem (ES) cells and compare their gene expression with that of primary PGC cultures. R1 (Nagy A et al. 1990 Development 110, 815–821) and Oct4-GiP (Ying QL et al. 2002 Nature 416, 545–548) ES cell lines were differentiated into PGCs. For in vitro differentiation, the modified method of Geijsen N et al. (2004 Nature 427, 148–154) was used. In brief, ES cell suspension was put into hanging drops (400 cells per drop) for two days, where they formed embryoid bodies (EBs). The medium consisted of Iscove's Modified Dulbecco's Media (Gibco) supplemented with 10% FBS (Hyclone, Logan, UT, USA), 30 μg mL−1 iron saturated transferrin (Gibco), 1 mM sodium pyruvate (Gibco), 0.1 mM 2-mercaptoethanol (Sigma, Hungary), non-essential amino acids (Sigma), 4.5 mM monothioglycerol (Sigma), 50 μg mL−1 ascorbic acid (Sigma), 2 mM glutamine (Gibco), and antibiotics. The EB clumps were differentiated in suspension culture for 2 or 5 days, and then dissociated with collagenase treatment. Cells positive for SSEA-1 were isolated from dissociated EBs by immunomagnetic bead sorting and plated into gelatinized plates in the presence of 2 μM retinoic acid (Sigma). After 7 days of culture, individual PGC colonies were isolated and subcloned. The subcloned PGCs were cultured in PGC medium consisting of Dulbecco's Modified Eagle Media (Gibco) supplemented with 15% FBS, 2 mM glutamine, 1 mM sodium pyruvate, 1000 U recombinant mouse leukaemia inhibitor factor (ESGRO®, Chemicon International, Inc., Temecula, CA, USA), 20 ng mL−1 basic fibroblast growth factor (Sigma), 60 ng mL−1 soluble mouse stem cell factor (Sigma), and antibiotics. The gene expression profile was monitored using semi-quantitative RT-PCR. The gene expression of Oct4, Nanog, Stella, Piwil2, Rnf17, and Tex14 were analyzed during the differentiation. Primary PGC cultures were also isolated from (C57BL/6 × DBA)F1 embryos of age 8.5 and 11.5 days post-coitum, and differentiated in vitro. The previously described PGC medium was used to proliferate the isolated cells. The gene expression profile of PGCs and ES-derived PGC lines were compared. There were no great differences between the gene expression profiles of PGCs and ES cell-derived PGC cells. SSEA-1 and alkaline phosphatase staining of cells did not show differences between the two cell populations. We have shown here the two PGC populations do not differ from each other in gene expression of the selected genes. Further investigation is needed to differentiate the PGCs into gametes and to analyze the gene expression of other genes involved in gamete differentiation. The authors would like to acknowledge Gyorgyi Kungl for the technical help. This research was supported by OTKA T046171 grant.

scholarly journals Gene Expression Profile of the Human Colorectal Carcinoma LoVo Cells Treated With Sporamin and Thapsigargin

2021 ◽  
Vol 11 ◽  
Author(s):  
Chun Yang ◽  
Si-Jia Chen ◽  
Bo-Wen Chen ◽  
Kai-Wen Zhang ◽  
Jing-Jie Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Ipomoea Batatas ◽  
Expression Profiles ◽  
Rna Seq ◽  
Ppi Network ◽  
Illumina Platform

Sporamin, a proteinase inhibitor isolated from the sweet potato (Ipomoea batatas), has shown promising anticancer effect against colorectal cancer (CRC) in vitro and in vivo but its mechanisms of action are poorly understood. In the present study, high throughput RNA sequencing (RNA-seq) technology was applied to explore the transcriptomic changes induced by sporamin in the presence of thapsigargin (TG), a non-12-O-tetradecanolphorbol-13-acetate type cancer promoter, in the LoVo human CRC cells. Cellular total RNA was extracted from the cells after they were treated with vehicle (CTL), 1 μM of thapsigargin (TG), or 1 μM of TG plus 30 μM of sporamin (TGSP) for 24 h. The migratory capacity of the cells was determined by wound healing assay. The gene expression profiles of the cells were determined by RNA-seq on an Illumina platform. GO enrichment analysis, KEGG pathway analysis, protein-protein interaction (PPI) network construction, and transcription factors (TF) prediction were all performed based on the differentially expressed genes (DEGs) across groups with a series of bioinformatics tools. Finally, the effect and potential molecular targets of the sporamin at the transcriptome level were evaluated. Sporamin significantly inhibited the migration of cells induced by TG. Among the 17915 genes detected in RNA-seq, 46 DEGs were attributable to the effect of sporamin. RT-PCR experiment validated that the expression of RGPD2, SULT1A3, and BIVM-ERCC5 were up-regulated while NYP4R, FOXN1, PAK6, and CEACAM20 were down-regulated. Sporamin enhanced the mineral absorption pathway, worm longevity regulating pathway, and pyrimidine metabolism pathway. Two TFs (SMIM11A and ATOH8) were down-regulated by sporamin. HMOX1 (up-regulated) and NME1-NME2 (down-regulated) were the main nodes in a PPI network consisting of 16 DEGs that were modulated by sporamin in the presence of TG. Sporamin could favorably alter the gene expression profile of CRC cells, up-regulating the genes that contribute to the homeostasis of intracellular metal ions and the activities of essential enzymes and DNA damage repairment. More studies are warranted to verify its effect on specific genes and delineate the mechanism of action implicated in the process.

scholarly journals Effect of Chronic Valproic Acid Treatment on Hepatic Gene Expression Profile inWfs1Knockout Mouse

PPAR Research ◽  
2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Marite Punapart ◽  
Mall Eltermaa ◽  
Julia Oflijan ◽  
Silva Sütt ◽  
Anne Must ◽  
...  
Keyword(s):  
Gene Expression ◽  
Valproic Acid ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Expression Profiles ◽  
Peroxisome Proliferator ◽  
Hepatic Gene Expression ◽  
Wild Type ◽  
Drug Induced

Valproic acid (VPA) is a widely used anticonvulsant and mood-stabilizing drug whose use is often associated with drug-induced weight gain. Treatment with VPA has been shown to upregulateWfs1expressionin vitro. Aim of the present study was to compare the effect of chronic VPA treatment in wild type (WT) andWfs1knockout (KO) mice on hepatic gene expression profile. Wild type,Wfs1heterozygous, and homozygous mice were treated with VPA for three months (300 mg/kg i.p. daily) and gene expression profiles in liver were evaluated using Affymetrix Mouse GeneChip 1.0 ST array. We identified 42 genes affected byWfs1genotype, 10 genes regulated by VPA treatment, and 9 genes whose regulation by VPA was dependent on genotype. Among the genes that were regulated differentially by VPA depending on genotype was peroxisome proliferator-activated receptor delta (Ppard), whose expression was upregulated in response to VPA treatment in WT, but not inWfs1KO mice. Thus, regulation ofPpardby VPA is dependent onWfs1genotype.

Gene expression profile in monocyte during in vitro mineral fiber degradation

2007 ◽  
Vol 82 (6) ◽  
pp. 355-362 ◽  
Author(s):  
Hermine Dika Nguea ◽  
Aymon de Reydellet ◽  
Patrice Lehuédé ◽  
Alain De Meringo ◽  
Alain Le Faou ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Mineral Fiber ◽  
Fiber Degradation

The t(14;18) defines a unique subset of diffuse large B-cell lymphoma with a germinal center B-cell gene expression profile

Blood ◽  
2002 ◽  
Vol 99 (7) ◽  
pp. 2285-2290 ◽  
Author(s):  
James Z. Huang ◽  
Warren G. Sanger ◽  
Timothy C. Greiner ◽  
Louis M. Staudt ◽  
Dennis D. Weisenburger ◽  
...  
Keyword(s):  
Gene Expression ◽  
B Cell ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Germinal Center ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Germinal Center B Cell ◽  
Cell Gene Expression ◽  
Cell Gene

Recently we have identified subgroups of de novo primary diffuse large B-cell lymphoma (DLBCL) based on complementary DNA microarray-generated gene expression profiles. To correlate the gene expression profiles with cytogenetic abnormalities in these DLBCLs, we examined the occurrence of the t(14;18)(q32;q21) in the 2 distinctive subgroups of DLBCL: one with the germinal center B-cell gene expression signature and the other with the activated B cell–like gene expression signature. The t(14;18) was detected in 7 of 35 cases (20%). All 7 t(14;18)-positive cases had a germinal center B-cell gene expression profile, representing 35% of the cases in this subgroup, and 6 of these 7 cases had very similar gene expression profiles. The expression of bcl-2 and bcl-6 proteins was not significantly different between the t(14;18)-positive and -negative cases, whereas CD10 was detected only in the group with the germinal center B-cell expression profile, and CD10 was most frequently expressed in the t(14;18)-positive cases. This study supports the validity of subdividing DLBCL into 2 major subgroups by gene expression profiling, with the t(14;18) being an important event in the pathogenesis of a subset of DLBCL arising from germinal center B cells. CD10 protein expression is useful in identifying cases of DLBCL with a germinal center B-cell gene expression profile and is often expressed in cases with the t(14;18).

scholarly journals Reversible programming of pluripotent cell differentiation

2000 ◽  
Vol 113 (3) ◽  
pp. 555-566 ◽  
Author(s):  
J. Lake ◽  
J. Rathjen ◽  
J. Remiszewski ◽  
P.D. Rathjen
Keyword(s):  
Gene Expression ◽  
Embryoid Bodies ◽  
Visceral Endoderm ◽  
Es Cell ◽  
Primitive Endoderm ◽  
Pluripotent Cell ◽  
Pluripotent Cells ◽  
Differentiation Potential

We have undertaken an in vitro differentiation analysis of two related, interconvertible, pluripotent cell populations, ES and early primitive ectoderm-like (EPL) cells, which are most similar in morphology, gene expression, cytokine responsiveness and differentiation potential in vivo to ICM and early primitive ectoderm, respectively. Pluripotent cells were differentiated in vitro as aggregates (embryoid bodies) and the appearance and abundance of cell lineages were assessed by morphology and gene expression. Differentiation in EPL cell embryoid bodies recapitulated normal developmental progression in vivo, but was advanced in comparison to ES cell embryoid bodies, with the rapid establishment of late primitive ectoderm specific gene expression, and subsequent loss of pluripotent cell markers. Nascent mesoderm was formed earlier and more extensively in EPL cell embryoid bodies, and resulted in the appearance of terminally differentiated mesodermal cell types prior to and at higher levels than in ES cell embryoid bodies. Nascent mesoderm in EPL cell embryoid bodies was not specified but could be programmed to alternative fates by the addition of exogenous factors. EPL cells remained competent to form primitive endoderm even though this is not the normal fate of primitive ectoderm in vivo. The establishment of primitive ectoderm-like gene expression and inability to participate in embryogenesis following blastocyst injection is therefore not directly associated with restriction in the ability to form extra-embryonic lineages. However, the EPL cell embryoid body environment did not support differentiation of primitive endoderm to visceral endoderm, indicating the lack of an inductive signal for visceral endoderm formation deduced to originate from the pluripotent cells. Similarly, the inability of EPL cells to form neurons when differentiated as embryoid bodies was attributable to perturbation of the differentiation environment and loss of inductive signals rather than a restricted differentiation potential. Reversion of EPL cells to ES cells was accompanied by restoration of ES cell-like differentiation potential. These results demonstrate the ability of pluripotent cells to adopt developmentally distinct, stable cell states with altered differentiation potentials.

scholarly journals Gene Expression Profiles of Papillary Thyroid Microcarcinoma

2017 ◽  
Vol 102 (1-2) ◽  
pp. 39-46 ◽  
Author(s):  
Woo Young Kim ◽  
Jae Bok Lee ◽  
Seung Pil Jung ◽  
Hoon Yub Kim ◽  
Sang Uk Woo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Profile ◽  
Differentially Expressed Genes ◽  
Expression Profile ◽  
Expression Profiles ◽  
Differentially Expressed ◽  
Papillary Thyroid Microcarcinoma ◽  
Papillary Thyroid ◽  
Normal Thyroid ◽  
Thyroid Microcarcinoma

The objective was to identify gene expression profile of papillary thyroid microcarcinoma. To help improve diagnosis of papillary thyroid microcarcinoma, we performed gene expression profiling and compared it to pair normal thyroid tissues. We performed microarray analysis with 6 papillary thyroid microcarcinoma and 6 pair normal thyroid tissues. Differentially expressed genes were selected using paired t test, linear models for microarray data, and significance analysis of microarrays. Real-time quantitative reverse transcription–polymerase chain reaction was used to validate the representative 10 genes (MET, TIMP1, QPCT, PROS1, LRP4, SDC4, CITED1, DPP4, LRRK2, RUNX2). We identified 91 differentially expressed genes (84 upregulated and 7 downregulated) in the gene expression profile and validated 10 genes of the profile. We identified a significant genetic difference between papillary thyroid microcarcinoma and normal tissue by 10 upregulated genes greater than 2-fold (P < 0.05).

scholarly journals Expression of Genes Related to Germ Cell Lineage and Pluripotency in Single Cells and Colonies of Human Adult Germ Stem Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-17 ◽  
Author(s):  
Sabine Conrad ◽  
Hossein Azizi ◽  
Maryam Hatami ◽  
Mikael Kubista ◽  
Michael Bonin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Adult Stem Cells ◽  
Single Cells ◽  
Human Fibroblasts ◽  
Embryonic Stem ◽  
Human Adult

The aim of this study was to elucidate the molecular status of single human adult germ stem cells (haGSCs) and haGSC colonies, which spontaneously developed from the CD49f MACS and matrix- (collagen−/laminin+ binding-) selected fraction of enriched spermatogonia. Single-cell transcriptional profiling by Fluidigm BioMark system of a long-term cultured haGSCs cluster in comparison to human embryonic stem cells (hESCs) and human fibroblasts (hFibs) revealed that haGSCs showed a characteristic germ- and pluripotency-associated gene expression profile with some similarities to hESCs and with a significant distinction from somatic hFibs. Genome-wide comparisons with microarray analysis confirmed that different haGSC colonies exhibited gene expression heterogeneity with more or less pluripotency. The results of this study confirm that haGSCs are adult stem cells with a specific molecular gene expression profilein vitro, related but not identical to true pluripotent stem cells. Under ES-cell conditions haGSC colonies could be selected and maintained in a partial pluripotent state at the molecular level, which may be related to their cell plasticity and potential to differentiate into cells of all germ layers.

Novel Murine Model To Study Modulation of Genes and Molecular Pathways Induced Following In Vivo Interaction between Multiple Myeloma Cells and Human BM Milieu.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3409-3409
Author(s):  
Paola Neri ◽  
Pierfrancesco Tassone ◽  
Masood Shammas ◽  
Mariateresa Fulciniti ◽  
Yu-Tzu Tai ◽  
...  
Keyword(s):  
Gene Expression ◽  
Multiple Myeloma ◽  
Bone Marrow ◽  
Cell Growth ◽  
Gene Expression Profile ◽  
Expression Profile ◽  
Murine Model ◽  
Cell Signalling

Abstract Interaction between multiple myeloma (MM) cells and the bone marrow (BM) microenvironment plays a critical role in promoting MM cell growth, survival, migration and development of drug resistance. This interaction within the bone marrow milieu is unique and its understanding is important in evaluating effects of novel agents in vitro and in vivo. We here describe a novel murine model that allows us to study the expression changes in vivo in MM cells within the human BM milieu. In this model, the green fluorescent protein (INA-6 GFP+) transduced IL-6-dependent human MM cell line, INA-6, was injected in human bone chip implanted into SCID mice. At different time points the bone chip was retrieved, cells flushed out and GFP+ MM cells were purified by CD138 MACS microbeads. Similar isolation process was used on INA-6 GFP+ cells cultured in vitro and used as control. Total RNA was isolated from these cells and gene expression profile analyzed using the HG-U133 array chip (Affymetrix) and DChip analyzer program. We have identified significant changes in expression of several genes following in vivo interaction between INA-6 and the BM microenvironment. Specifically, we observed up-regulation of genes associated with cytokines (IL-4, IL-8, IGFB 2–5) and chemokines (CCL2, 5, 6, 18, 24, CCR1, 2, 4), implicated in cell-cell signalling. Moreover genes implicated in DNA transcription (V-Fos, V-Jun, V-kit), adhesion (Integrin alpha 2b, 7, cadherin 1 and 11) and cell growth (CDC14, Cyclin G2, ADRA1A) were also up-regulated and genes involved in apoptosis and cell death (p-57, BCL2, TNF1a) were down-regulated. Using the Ingenuity Pathway Analysis the most relevant pathways modulated by the in vivo interaction between MM cells and BMSCs were IL-6, IGF1, TGF-beta and ERK/MAPK-mediated pathways as well as cell-cycle regulation and chemokine signalling. These results are consistent with previously observed in vitro cell signalling studies. Taken together these results highlight the ability of BM microenvironment to modulate the gene expression profile of the MM cells and our ability to in vivo monitor the changes. This model thus provides us with an ability to study in vivo effects of novel agents on expression profile of MM cells in BM milieu, to pre-clinically characterize their activity.

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