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Transcript abundance in A-T-derived iPSC: Comparing isogenic cells to unrelated individual

ABSTRACT: Induced pluripotent stem cells (iPSC) were prepared from multiple subjects with Ataxia-telangiectasia (A-T). iPSC were prepared from activated T-cells using commercial Sendai virus to deliver reprogramming factors. ATM protein-expressing and non-expressing cultures would found in multiple sub-lines (sub-lines are generated from distinct founder colonies of cells) from a single subject (Q3). Genetic analysis determined that all sub-lines originated from the same subject and were likely the product of spontaneous reversion by gene correction. To compare gene expression profiles, three iPSC samples were assayed: (1) A reverted ATM+/- subline from subject Q3, (2) An ATM-/- subline from subject Q3, and (3) An iPSC line from an unrelated ATM-/- subject, Q1. Analysis reveals that the majority of significantly-different genes between the unrelated ATM-/- line (Q1SA) and the reverted ATM+/- line (Q3SC) was due to genetic variation between individuals. A more focused set of contrasting genes could be identified between the isogenic Q3-derived lines (Q3SA, ATM-/-, vs. Q3SC, ATM+/-). The 206 transcripts that are significantly different point to a differential regulation of p53-associated pathways.

ORGANISM(S): Homo sapiens

PROVIDER: GSE71878 | GEO | 2015/11/19

SECONDARY ACCESSION(S): PRJNA292390

REPOSITORIES: GEO

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Transcript abundance in A-T-derived iPSC: Comparing isogenic cells to unrelated individual

Project description:Induced pluripotent stem cells (iPSC) were prepared from multiple subjects with Ataxia-telangiectasia (A-T). iPSC were prepared from activated T-cells using commercial Sendai virus to deliver reprogramming factors. ATM protein-expressing and non-expressing cultures would found in multiple sub-lines (sub-lines are generated from distinct founder colonies of cells) from a single subject (Q3). Genetic analysis determined that all sub-lines originated from the same subject and were likely the product of spontaneous reversion by gene correction. To compare gene expression profiles, three iPSC samples were assayed: (1) A reverted ATM+/- subline from subject Q3, (2) An ATM-/- subline from subject Q3, and (3) An iPSC line from an unrelated ATM-/- subject, Q1. Analysis reveals that the majority of significantly-different genes between the unrelated ATM-/- line (Q1SA) and the reverted ATM+/- line (Q3SC) was due to genetic variation between individuals. A more focused set of contrasting genes could be identified between the isogenic Q3-derived lines (Q3SA, ATM-/-, vs. Q3SC, ATM+/-). The 206 transcripts that are significantly different point to a differential regulation of p53-associated pathways. Total cellular RNA was prepared from each iPSC culture. Different passage numbers or sister cultures were used as replicates (n=2).

2015-11-19 | E-GEOD-71878 | biostudies-arrayexpress

RNA-Sequencing of mouse melanoma cell line WT31 and its subline WT31_Passage

Project description:RNA-Sequencing analysis of in vitro cultured, sub-confluent (50-75%) mouse melanoma cell line WT31 and its subline WT31_Passage. The subline was generated by repeated in vivo hepatic passaging (5 times).

2023-05-11 | GSE219236 | GEO

DNA microarrays of 4 human iPSC cell lines and the ESC cell line H9

Project description:Documents of DNA expression of 4 human induced pluripotent stem cell (iPSC) lines from umbilical cord mesenchymal cells (UMCs) and amniotic mesenchymal cells (AMCs). We used microarrays to identify similarity between 4 iPSC cell lines and the human embryonic stem cell (ESC) line H9. 2 AMC iPSC cell lines, 2 UMC iPSC cell lines, H9 ESC cell line. TRIZOL cell lysates were prepared.

2010-11-10 | E-GEOD-20581 | biostudies-arrayexpress

MicroRNAome of highly metastatic and non-metastatic human breast cell lines

Project description:MicroRNAs are noncoding, endogenous small RNAs that regulate target genes by cleavage of the targeted mRNA or translational repression. We investigated the microRNAome using 2-color microarrays in a highly invasive human breast cancer cell line, MDA-MB-231 (sub line 4175) and a non-invasive breast epithelial cell line, MCF10A. We found 13 miRNAs that were up-regulated and 9 were down-regulated significantly in 4175 cells (p <0.05, fold change >2) compared with MCF10A cells. We compared the highly metastatic human breast cell lines MDA-MB-231 (4175 subline) with non-metastatic MCF10A cell lines. Two 4175 sublines and two MCF10A cell lines, independently grown and harvested. Dye swap was performed.

2010-05-15 | E-GEOD-17350 | biostudies-arrayexpress

Maturation,not initiation, is major roadblock during reprogramming toward pluripotency from human fibrobalsts

Project description:Reprogrammed cells are reverted to non reprogrammed cell fate in a maturation step of reprogramming. Cell fate changes during reprogramming from fibroblasts to iPSC , N=60 fibrobalst lines, N=13 Undifferentiated pluripotent stem cells, N=16 Nascent reprogrammed cells, N=31

2013-07-01 | E-GEOD-47489 | biostudies-arrayexpress

Multi-omics data and stochastic simulations support a cancer heterogeneity framework unifying genetic, epigenetic, and stochastic variabilities [Bulk-RNA-seq]

Project description:We subject multiple versions and single cell-derived sublines of an archetypal non-small cell lung cancer cell line to experimental scrutiny at the genomic, transcriptomic, and cell population levels. We find evidence of genetic variability among cell line versions and one subline. Additional sublines show no genetic variation but epigenetic differences are detected at the transcriptomic level. Stochastic simulations of subline growth dynamics confirm that one subline likely contains at least two genetic states while all others are isogenic. Overall, our results substantiate a view of intratumoral heterogeneity in which different genetic states give rise to multiple epigenetic “basins of attraction,” across which cells can transition driven by stochastic factors such as gene expression noise and asymmetric cell division. Deconvolving intratumoral heterogeneity into genetic, epigenetic, and stochastic components provides a lens through which to view tumor drug response and acquired treatment resistance that may lead to novel therapeutic strategies in the future.

2020-05-08 | GSE150025 | GEO

Multi-omics data and stochastic simulations support a cancer heterogeneity framework unifying genetic, epigenetic, and stochastic variabilities [scRNA-seq]

2020-05-08 | GSE150083 | GEO

Gene expression profiles new methotrexate resistant cell lines for the study of gestational trophoblastic neoplasia

Project description:Two trophoblastic cell lines, CRL-1584 (3A-Sub E) and JEG-3 were purchased from American Type Culture collection (ATCC) (Manassas, VA). CRL-1584 (3A-Sub E) was originally derived from human term primary placental cells and then immortalized using SV40 virus. JEG-3 is a clonal human choriocarcinoma. The cell lines were serially passaged in complete medium supplemented with each cell lineâs respective IC50 concentration of methotrexate until the cell lines were able to proliferate normally. Subsequently, the concentration of methotrexate was increased by one half-log concentration. The process was repeated iteratively until the cells became senescent. At each log concentration of methotrexate resistance, cell line stocks were frozen to establish a separate subline of each cell line. Based on their relative level of resistance to methotrexate, these were designated JEG-3-R7, JEG-3-R6, and JEG-3-R5 (for 10-7 M through 10-5 M methotrexate resistance). Over the same time period (approximately 11 months), we were able to induce only one order of magnitude of resistance in the normal placenta cell line; that line was designated NP-R7. In parallel, the parent cell lines were passaged in normal medium to control for any physiologic changes induced by prolonged culture in vitro.

2017-10-17 | E-MTAB-5185 | biostudies-arrayexpress

Enhancement of metastasis by spontaneous ploidy doubling of breast cancer cell MDA-MB-231.

Project description:MDA-MB-231 bone-metastatic subline 1833 and lung metastatic subline 4175 underwent spontaneous ploidy doubling in culture, i.e. the genome approximately duplicated itself gradually. The modal- and hyper-ploid subpopulations during the ploidy transition were sorted into two separate sublines, 1833-Modal and 1833-Hyper for 1833, 4175-Modal and 4175-Hyper for 4175. Their expresssion patterns were compared to each other as well as to other MDA-MB-231 sublines isolated previously by Kang et al. 2003 and Minn et al. 2005. Keywords: Cell type comparison 19 cell lines were analyzed, including the parental line MDA-MB-231, modal-ploid sublines 1833-Modal and 4175-Modal, hyper-ploid sublines 1833-Hyper and 4175-Hyper, strongly bone-metastatic lines 1833 (the original subline after short culture), 2274, 2268 and 2269, weakly bone-metastatic lines 2293, 2295 and 2297, strongly lung-metastaic lines 4142, 4173, 4175 (the original subline after short culture) and 4180, and weakly lung-metastatic lines SCP6, SCP21 and SCP26. Single sample for each line.

2010-06-08 | E-GEOD-16554 | biostudies-arrayexpress

Coding mutations and loss-of-imprinting in human pluripotent cells derived by nuclear transfer and defined factors [RNA-Seq]

Project description:Human pluripotent stem cells can be derived from somatic cells by forced expression of defined factors, and more recently by nuclear-transfer into human oocytes, revitalizing a debate on whether one reprogramming approach might be advantageous over the other. Here we compared the genetic and epigenetic stability of human nuclear-transfer embryonic stem cell (NT-ESC) lines and isogenic induced pluripotent stem cell (iPSC) lines, derived from the same somatic cell cultures of fetal, neonatal and adult origin. Both cell types shared similar genome-wide gene expression and DNA methylation profiles. Importantly, NT-ESCs and iPSCs have comparable numbers of de novo coding mutations but significantly higher than parthenogenetic ESCs. Similar to iPSCs NT-ESCs displayed clone- and gene-specific aberrations in DNA methylation and allele-specific expression of imprinted genes, similarly to iPSCs. The occurrence of these genetic and epigenetic defects in both NT-ESCs and iPSCs suggests that they are inherent to reprogramming, regardless of the underlying technique. RNA sequencing analysis was performed on a total of 12 human cell lines, including: an isogenic set of 3 nuclear-transfer embryonic stem cell (NT-ESC) lines, 2 RNA-reprogrammed induced pluripotent stem cell (iPSC) lines and their parental neonatal fibroblast cell line; an isogenic set of 1 NT-ESC line, 3 iPSC lines and their parental adult fibroblast cell line (derived from a type 1 diabetic subject); as well as 1 control embryonic stem cell (ESC) line.

2014-11-06 | E-GEOD-61656 | biostudies-arrayexpress

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