Comparison of the gene expression profiles between iPS-OSH, iPS-J, ESC and MEF.
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ABSTRACT: iPS-OSH was the novel iPS cells generated from our lab, the microarray analysis was used for determining the gene expression profiles of iPS-OSH. iPS-J was provided from Dr. Yamanaka (Riken, APS0001). Microarray analysis was used to compare the difference of gene expression profiles in iPS-OSH, iPS-J, ESC and MEF. The total RNA extracted from iPS-OSH, iPS-J, ESC and MEF was used for the microarray analysis to compare the gene expression profiles.
Project description:The microarray analysis was used for comparing the gene expression profiles between MEF, MEF treated with n-Butylenephthalide (BP) 10 and 40ug/ml. The total RNA extracted from MEF, MEF treated with n-Butylenephthalide (BP) 10 and 40ug/ml was used for the microarray analysis to compare the gene expression profiles.
Project description:Induced pluripotent stem (iPS) cells were produced from reprogramming of somatic cells, and they are shown to possess pluripotent properties similar to embryonic stem (ES) cells. Here we used microarrays to detail the global expression pattern among the ES cells and iPS cells, as well as the original mouse embryo fibroblast (MEF), to identify important players involved in the reprogramming process. Experiment Overall Design: Mouse ES cell cultures, as well as selected iPS cell lines and the original MEF cells they were derived from, were used for RNA extraction and hybridization on Affymetrix microarrays. Three biological replicates for each sample were processed. GeneChips were processed and data were analyzed as previously described (Zeng et al., Dev Biol 20004).
Project description:Induced pluripotent stem (iPS) cells were produced from reprogramming of somatic cells, and they are shown to possess pluripotent properties similar to embryonic stem (ES) cells. Here we used microarrays to detail the global expression pattern among the ES cells and iPS cells, as well as the original mouse embryo fibroblast (MEF), to identify important players involved in the reprogramming process. Mouse ES cell cultures, as well as selected iPS cell lines and the original MEF cells they were derived from, were used for RNA extraction and hybridization on Affymetrix microarrays. Three biological replicates for each sample were processed. GeneChips were processed and data were analyzed using the RMA package of Bioconductor.
Project description:The generation of induced pluripotent stem cells (iPSCs) often results in aberrant silencing of the imprinted Dlk1-Dio3 gene cluster, which compromises their ability to generate entirely iPSC-derived mice (âall-iPSC miceâ). Here, we show that reprogramming in the presence of ascorbic acid attenuates hypermethylation of Dlk1-Dio3 by enabling a chromatin configuration at its imprint control region that interferes with abnormal binding of the DNA methyltransferase Dnmt3a. This approach allowed us to generate adult all-iPSC mice from mature B cells, which have thus far failed to support the development of exclusively iPSC-derived postnatal mice. Our data demonstrate that factor-mediated reprogramming can endow a defined, terminally differentiated cell type with a developmental potential equivalent to that of embryonic stem cells. More generally, these findings indicate that the choice of culture conditions used for transcription factor-mediated reprogramming can strongly influence the epigenetic and biological properties of resultant iPSCs. This series consists of quadruplicated mRNA expression microarray data (Affymetrix mouse 430_2 3'-IVT array) for iPS cells derived from MEF cells under cell culture conditions with or without ascorbic acid supplementation. iPS cells were generated from MEFs of the Col-OKSM reprogrammable mice. In the presence of doxycycline, the reprogramming transcription factors Oct4, Sox2, Klf4, and cMyc were induced in MEFs to derivate iPS cells. Total RNA was isolated from iPS cells derivated in the presence or absence of ascorbic acid in culture medium.
Project description:It has been shown that DNA demethylation has a pivotal role in the generation of induced pluripotent stem (iPS) cells. However, the underlying mechanism is still unclear. Previous reports indicated that activation-induced cytidine deaminase (Aid) is involved in DNA demethylation in several developmental processes and cell fusion-mediated reprogramming. Based on the reports, we hypothesized that Aid may be involved in DNA demethylation during the iPS cell generation. In this study, we examined the function of Aid in iPS cell generation using Aid knockout (Aid-/-) mice expressing a GFP reporter under the control of a pluripotent stem cell marker, Nanog. By the introduction of Oct3/4, Sox2, Klf4 and c-Myc, Nanog-GFP positive iPS cells could be generated from the fibroblasts and primary B cells of Aid-/- mice. The Aid-/- iPS cells showed normal proliferation and gave rise to chimeras, indicating their capacity for self-renewal and pluripotency. Microarray analysis demonstrated that the global gene expression of Aid-/- iPS cells was similar to that of Aid+/+ iPS cells. Aid+/+ and Aid-/- iPS colonies were generated from Aid+/+ and Aid-/- MEFs and picked up mechanically. The clones were passaged four times on feeder cells and two times on gelatin-coated dishes to exclude the contamination of feeder cells. Subsequently, the RNA was isolated. Six Aid+/+ iPS cell clones and six Aid-/- iPS cell clones were compared by microarray. Samples from Aid+/+ and Aid-/-iPS cells
Project description:Transcriptional profiling of Mouse iPS cells and MEFs comparing ES cells. RNA was prepared from the ES cell line CGR8.8, MEFs and and iPSC clone and cRNA was hybridized to Agilent Whole Murine Genome Oligo Microarray.
Project description:iPS-OSH was the novel iPS cells generated from our lab, the microarray analysis was used for determining the gene expression profiles of iPS-OSH. iPS-J was provided from Dr. Yamanaka (Riken, APS0001). Microarray analysis was used to compare the difference of gene expression profiles in iPS-OSH, iPS-J, ESC and MEF.
Project description:We were able to achieve an initial stable intermediate phase by the transduction of Oct4, Klf4, and c-Myc. Furthermore, over-expression of Sox2 in these intermediate stage cells leads to final iPS cell phase. After examining the gene expression profiles from the initial to final iPS cell phases, we have identified Sox2 downstream genes important for iPS cell induction. 4 groups of cells are analyzed. Each group contains cells from 4 indepent dishes. The four groups are: ES cells, iPS cells, intermediate stage cells (OKM cells) and MEFs
Project description:To realize the gene expression in response to acute heat stress in chicken testis, we have employed whole genome microarray expression profiling as we have employed whole genome microarray expression profiling as a tool to identify genes response to acute heat stress. Male B strain Taiwan country chickens were subjected to acute heat stress (38M-bM-^DM-^C) for 4 h, and then exposed to 25M-bM-^DM-^C, with testes collected 0, 2, and 6 h after the cessation of heat stress, using non heat-stressed roosters as a control group (n = 3 roosters per group). Based on a chicken 44K oligo microarray, 163 genes significantly differed in the testes of the heat-stressed chickens from those of the control chickens. The mRNA expressions of upregulated genes, including HSP25, HSP90AA1, HSPA2, and LPAR2, and downregulated genes, including CDH5, CTNNA3, EHF, CIRBP, SLA, and NTF3, were confirmed through quantitative real-time polymerase chain reaction (qRT-PCR). Acute heat stress induced testicular gene experssion in B strain Taiwan country chicken was measured at 0, 2, and 6 h of recovery after 4 h of 38 degree acute heat stress.
Project description:Using magneto-nanofection, iPS cells were efficiently generated by the transient expression of iPS genes in MEF (mouse embryonic fibroblast), suggesting that the non-viral magneto-nanofection method can be used for the efficient generation of iPS cells.