Expression data from human osteosarcoma bone primaries or lung metastases
ABSTRACT: The high concentration of Well5 cells was resuspended into 20μl PBS, the needle along the tibia direction, before reaching in a breakthrough sense, direct injection cells. At 7 days after injection, proximal tibia was able to reach mass production. At 20 days after injection, the proximal tibia mass increased.If prolonging exposure by BLI,this stage displayedthat tumor cell signalsbegan to lung metastasis. Osteosarcoma orthotopic lung metastasis model was successfully constructed. Total RNA was extracted from sorted osteosarcoma cells of the primary site and lung metastases using Trizol (Invitrogen). We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during osteosarcoma lung metastasis. In support of the notion that fibrosis marks the lung metastasis, the expression of numerous fibrosis-related genes such as FN1, COLs, and MMPs were upregulated from the primary site to lung metastasis in Well5-luc orthotopic inoculation model. Total RNA was extracted from sorted osteosarcoma cells using Trizol (Invitrogen). Gene expression profiling was conducted by Shanghai Biotechnology Corporation using Affymetrix U133 plus 2.0 arrays (Affymetrix, Santa Clara, CA). All data were analyzed according to the manufacturer’s protocol. Raw data generated from Affymetrix CEL files were normalized by RMA background correction; values were log2 transformed. For the enrichment of P values of each GO term, we used Fisher’s exact test to calculate P values and R package stats to calculate FDR (q value) by BH method (www.r-project.org).
Project description:Osteosarcoma is thought of arising from the malignant transformation of osteogenic progenitors. The stage-specific embryonic antigen-4 (SSEA-4) labels a subset of TICs specially present in the high-risk subgroup. SSEA-4+ AND SSEA-4-
Project description:Lung ischemia-reperfusion (I/R) injury remains one of the common complications after various cardiopulmonary surgeries. I-R injury represents one potentially maladaptive response of the innate immune system which is featured by an exacerbated sterile inflammatory response triggered by tissue damage. Thus, understanding the key components and processes involved in sterile inflammation during lung I-R injury is critical to alter care and extend survival for patients with acute lung injury. We constructed a minipig surgical model of transient unilateral left pulmonary artery occlusion without bronchial involvement to create ventilated lung I-R injury. Lung tissues from minipig with sham operation (one sample), left side lung tissues (the operated side)(one sample) and right side lung tissues (the non-operated side)(one sample) from minipig with lung ischemia-reperfusion were submitted for gene expression array analysis.
Project description:Generation of haploid gametes in vitro can potentially address gamete failure-based infertility.This study reports complete in vitro meiosis from murine ESC-derived PGCLCs resulting in the formation of male spermatid-like cells (SLCs) capable of producing viable fertile offspring via intracytoplasmic sperm injection (ICSI).Our findings provide the basis for generation of haploid spermatids in vitro in human, the generation of transgenic animals, and the use of this system to investigate mechanisms of meiosis. We used microarrays to compare gene expression profiles of in vivo and in vitro derived PGC cells and round spermatids. We collected E12.5 male fatal PGCs, PGCLC in vitro, round spermatids and spermatids like cells produced in vitro, each sample has 3 replications.
Project description:Zygotic genome activation (ZGA), which is according to the midblastula transition in zebrafish, is an important event during the maternal-zygotic transition in animals. Our preliminary study and other group’s works indicate that epigenetic regulations play an essential role in ZGA. Morpholino was employed to knockdown PRMT6. We used microarrays to analyze the global gene expression in prmt6 morphants. prmt6 MO (0.3mM) was injected into the one-two cell zebrafish, prmt6 cMO (0.3mM) injection as a control. At 6 hpf, embryos were classified into three subtypes (normal, mild and severe) and prepared for global gene expression analysis with Affymetrix Zebrafish Genome Arrays. The severe subtype and the control were repeated three times.
Project description:Haploid stem cells offer an easy-to-manipulate genetic system and therefore have great values for studies of recessive phenotypes. Here, we show that mouse androgenetic haploid ES (ahES) cell lines can be established by transferring sperm into enucleated oocyte. The ahES cells maintain haploidy and stable growth over 30 passages, express pluripotent markers, possess the ability to differentiate into all three germ-layers in vitro and in vivo, and contribute to germline of chimeras when injected into blastocysts. Although epigenetically distinct from sperm cells, the ahES cells can produce viable and fertile progenies after intracytoplasmic injection into mature oocytes. The oocyte injection procedure can also produce viable transgenic mice from genetically engineered ahES cells. We used microarrays to compare the global programme of gene expression among ahES cells, normal diploid ES cells, MEF cells and round sperm cells and found that gene expression pattern of ahES cells was highly similar with ES cells but was distinct from MEF cells and round sperms. Androgenetic haploid ES cells were FACS sorted to harvest the G0/G1 phase haploid cells. Total RNA were extracted from three ahES cell lines (AH129-5, AH129-N1, AH129-NC1, all 129Sv genetic background), two ES cell lines ( CS1-1, R1, 129Sv background), MEF cells and round sperm and hybridized with Affymetrix GeneChip 430 2.0 array. Data were collected and analyzed to compare their gene expression pattern.
Project description:Haploid cells are amenable for genetic analysis because they contain only one set of chromosomes.Here,we report the derivation of haESCs from androgenetic blastocysts. These cells, which we designated AG-haESCs, express classical ESC markers, are pluripotent, and contribute to various tissues including the germline upon injection into diploid blastocysts. We used microarrays to compare the gene expression levels among androgenetic haploid embryonic stem cell lines(AG-haESC) E14 and male mouse embryonic fibroblasts (MEFs) and identified that most paternally imprinted genes were down-regulated and the maternally imprinted genes were up-regulated. To avoid the influence of diploidized cells on the expression profile, we collected samples from FACS of cells at G1/G0 stage by staining Hochest 33342. We used E14,which was a male embryonic stem cell lines, and MEFs isloated from male individuals as control. Gene expression profiles of all the cell lines were analysed on an Affymetrix GeneChip 430 2.0 array.
Project description:Compare difference Global expression profile of hiPSCs between hESCs and human Somatic cells, showing that hiPSCs and hESCs is consistent in lineages and indicated that the induce method is safe and reliable. There are three groups of samples, each group has two repeated samples, hiPSCs respectively compared with hESCs and human Urine-Derived Cells.
Project description:When PDMSCs were induced to heptocytes in vitro, cells mophology, stem cell markers, mitochondrial metabolism will change according to the differentiated status.But dedifferentiation reverses differentiated cells to a more primitive phenotype and PDMSCs will retain the multilineage potency. Furthermore, it will leads to the alteration of gene expression pattern. We used microarrays to detail the global programme of gene expression underlying dedifferentiation and hepatogenic differentiation prcocesses, we intend to identify distinct classes of differentiated genes during these processes. Human PDMSCs at passage 5 were induced to hepatocytes for 11 days, then the inductive medium was replaced by general culture medium for 1 day. Then human PDMSCs, hepatogenic PDMSCs at 11 days, dedifferentiated PDMSCs were selected for RNA extraction and hybridization on Affymetrix microarrays. To that end, we hand-selected cells at three time-points: before hepatogenic induction (P), hepatogenic PDMSCs at 11 days (H) and dedifferentiated PDMSCs for 1 day (DH) .
Project description:Long noncoding RNAs (lncRNAs) have been implicated in the formation of many different types of tumors. However, expression profiles and potential functions of lncRNAs in non-functioning pituitary adenomas (NFPAs) have not been systematically evaluated. We evaluated the expression profiles and potential functions of lncRNAs in non-functioning pituitary adenomas (NFPAs). 10 formalin-fixed and paraffin-embedded (FFPE) tissue specimens (5 non-functioning pituitary adenomas (NFPAs) and 5 normal pituitaries(NPs)) were selected for RNA extraction and hybridization on Affymetrix microarrays. The NFPAs team was designed as the Tumor group (T), while the NPs team was designed as Normal group (N).
Project description:Gliomas arising in the brainstem and thalamus are devastating tumors that are difficult to surgically resect due to their proximity to eloquent brain structures. Here, we performed a comprehesive genomic and epigenomic study, using gene expression and methylation microarrays, to research on th different genomic and epigenetic signatures between brainstem, thalamic, and supratentorial gliomas. Comparison of brainstem, thalamic and supratentorial gliomas