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Expression in irradiated MEFs exposed to murine acute lymphoblastic leukemia cells

ABSTRACT: Primary pre-B acute lymphoblastic (ALL) cells do not proliferate long-term ex vivo without the presence of stromal support. We developed and use an ex vivo co-culture model, consisting of mouse leukemic pre-B Bcr/Abl-expressing ALL cells grown with mitotically inactivated mouse embryonic fibroblasts (MEFs). This system provides a generic type of environmentally-mediated protection to the ALL cells, because when the ALL cells are treated with a moderate dose of a therapeutic drug, drug-resistant ALL cells can be recovered after a 1-2 week period of culture. Some of the factors produced by stromal cells that provide protection to ALL cells have been identified. However, it is unclear if the presence of drug-treated ALL cells affects the stromal fibroblasts. The current study was initiated to examine this using expression profiling on the irradiated MEFs.

ORGANISM(S): Mus musculus

PROVIDER: GSE33329 | GEO | 2011/12/01

SECONDARY ACCESSION(S): PRJNA148995

REPOSITORIES: GEO

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Expression in irradiated MEFs exposed to murine acute lymphoblastic leukemia cells

Project description:Primary pre-B acute lymphoblastic (ALL) cells do not proliferate long-term ex vivo without the presence of stromal support. We developed and use an ex vivo co-culture model, consisting of mouse leukemic pre-B Bcr/Abl-expressing ALL cells grown with mitotically inactivated mouse embryonic fibroblasts (MEFs). This system provides a generic type of environmentally-mediated protection to the ALL cells, because when the ALL cells are treated with a moderate dose of a therapeutic drug, drug-resistant ALL cells can be recovered after a 1-2 week period of culture. Some of the factors produced by stromal cells that provide protection to ALL cells have been identified. However, it is unclear if the presence of drug-treated ALL cells affects the stromal fibroblasts. The current study was initiated to examine this using expression profiling on the irradiated MEFs. We isolated RNA from irradiated wild-type MEFs after a 9-day culture period. MEFs had either been exposed to DMSO, to nilotinib, a drug that is not expected to affect them, or to ALL cells treated with DMSO or nilotinib. MEFs and ALL cells were physically separated by a Transwell membrane of a pore size that did not allow the ALL cells to migrate towards the MEFs, but allowed the exchange of diffusible molecules.

2011-12-01 | E-GEOD-33329 | biostudies-arrayexpress

Metabolic characterization of directly reprogrammed renal tubular epithelial cells (iRECs)

Project description:Fibroblasts can be directly reprogrammed to induced renal tubular epithelial cells (iRECs) using four transcription factors. These engineered cells may be used for disease modeling, cell replacement therapy or drug and toxicity testing. Direct reprogramming induces drastic changes in the transcriptional landscape, protein expression, morphological and functional properties of cells. However, how the metabolome is changed by reprogramming and to what degree it resembles the target cell type remains unknown. Using untargeted gas chromatography-mass spectrometry (GC-MS) and targeted liquid chromatography-MS, we characterized the metabolome of mouse embryonic fibroblasts (MEFs), iRECs, mIMCD-3 cells, and whole kidneys. Metabolic fingerprinting can distinguish each cell type reliably, revealing iRECs are most similar to mIMCD-3 cells and clearly separate from MEFs used for reprogramming. Treatment with the cytotoxic drug cisplatin induced typical changes in the metabolic profile of iRECs commonly occurring in acute renal injury. Interestingly, metabolites in the medium of iRECs, but not of mIMCD-3 cells or fibroblast could distinguish treated and non-treated cells by cluster analysis. In conclusion, direct reprogramming of fibroblasts into renal tubular epithelial cells strongly influences the metabolome of engineered cells, suggesting that metabolic profiling may aid in establishing iRECs as in vitro models for nephrotoxicity testing in the future.

2021-08-20 | MTBLS608 | MetaboLights

Gene expression data from the mesenchymal stem cell line OP9M2 that supported expansion of human hematopoietic stem and progenitor cells in vitro

Project description:One of the long-standing goals in the field has been to establish a culture system that would allow maintenance of HSC properties ex vivo. In the absence of such system, the ability to model human hematopoiesis in vitro has been limited, and there has been little progress in the expansion of human HSCs for clinical application. To that end, we defined a mesenchymal stem cell co-culture system based on a monoclonal OP9 stromal cell line (OP9M2), for expansion of clonally multipotent human HSPCs that were protected from apoptosis and immediate differentiation, and retained the HSPC phenotype. To identify the supportive mechanisms, we performed a genome-wide gene expression analysis of OP9M2 stromal cells and compared the expression to a non-supportive stomal line (BFC012). This co-culture system provides a new, well-defined platform for studying mechanisms involved in HSC-niche interactions and protection of critical HSC properties ex vivo. To determine the cellular identity and the supportive mechanism of the OP9M2 cells, we compared the OP9M2 cells with non-supportive BFC012 stromal cells using Affymetrix mouse microarrays.

2013-01-31 | E-GEOD-34973 | biostudies-arrayexpress

Environmental-mediated vincristine drug resistance of human precursor B-acute lymphoblastic leukemia (pre-B ALL)

Project description:First-line treatment for pre-B acute lymphoblastic leukemia (pre-B ALL) typically encompasses a period of so-called induction therapy in which the patient is treated for 28 days with chemotherapy to induce a remission. A bone marrow sample is subsequently taken to determine the number of remaining leukemia cells, which are called minimal residual disease (MDR). MRD is a key prognostic factor, with higher MRD levels predicting worse outcome for patients. Induction therapy usually is able to drastically reduce the number of leukemic cells but chemoresistance, as first detected by higher MRD levels, remains a major clinical problem. MRD persists not only by leukemia cell-intrinsic mechanisms such as clonal selection and genetic changes, but also through protective interactions of the leukemia cells with the surrounding supporting stroma. This mechanism of drug resistance has been named environment-mediated drug resistance (EMDR) and is a general route through which MRD cells, regardless of underlying genetics of the leukemia cells, can persist after chemotherapy. The EMDR component contributing to the persistence of MRD leukemia cells can be studied in tissue culture models by treating leukemia cells with a non-lethal drug dose in the presence of stromal cell protection. Here we analysed the transcriptome of a PDX-derived human pre-B ALL cell line co-cultured with OP9 stromal cells as they developed drug insensitivity against the chemotherapeutic drug vincristine.

2023-02-28 | GSE176366 | GEO

Expression data from porcine embryonic stem cells

Project description:We have been able to derive EpiSC-like pESC lines from in vivo produced porcine blastocysts. Our cell lines showed AP activity, expressions of the genes Oct4, Sox2, Nanog, Rex1, TDGF1, bFGF, FGFR1, FGFR2, Nodal and Activin-A involved in pluripotency and signaling pathways and in vitro differentiation potential, displaying similarities to epiblast stem cells or hES cells. Porcine blastocysts were cultured on mitotically inactivated mouse embryonic fibroblasts (MEFs) in pESC medium. Following 5-7 days of culture, we observed EpiSC-like primary colonies derived from day 7 in vivo-produced. These EpiSC-like pESC colonies were mechanically dissociated into several clumps using pulled glass pipettes 10-15 days after seeding. Dissociated clumps were then re-seeded on fresh MEFs, and subsequent EpiSC-like pESC lines were routinely passaged via the pulled glass pipette method every 5-7 days. Our cell lines maintained stemness and a stable morphology for more than 56 passages. The main purpose of the present study was to investigate gloval gene expression from porcine embryonic stem cells.

2011-12-31 | E-GEOD-32506 | biostudies-arrayexpress

Transcriptional Mediator Subunit MED1/TRAP220 in Stromal Cells Is Involved in Hematopoietic Stem/Progenitor Cell Support through Osteopontin Expression

Project description:MED1/TRAP220, a subunit of the transcriptional Mediator/TRAP complex, is crucial for various biological events through its interaction with distinct activators such as nuclear receptors and GATA family activators. In hematopoiesis, MED1 plays a pivotal role in optimal nuclear receptor-mediated myelomonopoiesis and GATA-1-induced erythropoiesis. In this study, we present evidence that MED1 in stromal cells is involved in supporting hematopoietic stem and/or progenitor cells (HSPCs) through osteopontin (OPN) expression. We found that the proliferation of bone marrow (BM) cells cocultured with MED1 knockout (Med1-/-) mouse embryonic fibroblasts (MEFs) was significantly suppressed when compared to the control. Furthermore, the number of long-term culture-initiating cells (LTC-ICs) was attenuated for BM cells cocultured with Med1-/- MEFs. The vitamin D receptor (VDR)- and Runx2-mediated expression of OPN, as well as Mediator recruitment to the Opn promoter, was specifically attenuated in the Med1-/- MEFs. Addition of OPN to these MEFs restored the growth of cocultured BM cells and the number of LTC-ICs, both of which were attenuated by the addition of the anti-OPN antibody to Med1+/+ MEFs and to BM stromal cells. Consequently, MED1 in niche appears to play an important role in supporting HSPCs, by upregulating VDR- and Runx2-mediated transcription on the Opn promoter. Total RNAs of mouse embryonic fibroblasts (MEFs) derived from Med1-/-/p53-/- and Med1+/+/p53-/- E10.0 embryos were compared. Duplicate samples for each MEF type were analyzed.

2010-08-31 | E-GEOD-22471 | biostudies-arrayexpress

Gene expression data from the mesenchymal stem cell line OP9M2 that supported expansion of human hematopoietic stem and progenitor cells in vitro

2013-01-31 | GSE34973 | GEO

Transcriptional Mediator Subunit MED1/TRAP220 in Stromal Cells Is Involved in Hematopoietic Stem/Progenitor Cell Support through Osteopontin Expression

2010-08-31 | GSE22471 | GEO

Expression profile of stromal cells in the lymph node

Project description:Tissue structure of the lymph node (LN) is supported by the network of stromal cells of mesenchymal origin, which is suggested to contribute to various immunological processes. In order to identify stromal cellM-bM-^@M-^Sderived factors that regulate immune function, we performed gene expression profiling of stromal cells freshly isolated from collagenase digested mouse LNs, cultured stromal cell lines, and embryonic fibroblasts as a mesenchymal control. Total RNAs extracted from the CD45-CD31-podoplanin+VCAM-1+ stromal cell fraction was sorted from enzymatically digested mouse LNs, cultured stromal cell lines stimulated with or without lymphotoxin (LTM-NM-2R agonistic antibody), and mouse embryonic fibroblasts (MEFs) were used for microarray analysis to assess relative gene expression.

2014-06-19 | E-GEOD-55251 | biostudies-arrayexpress

Gene expression profiles of epithelial and mesenchymal cells extracted from heterotypic ex vivo co-cultures or monocultures

Project description:Stromal-epithelial interactions play a fundamental role in tissue homeostasis, controlling cell proliferation and differentiation. Not surprisingly, aberrant stromal-epithelial interactions contribute to malignancies. The goals and objectives of this study were 1.) to characterize and validate the molecular identity of human primary epithelial and stromal/mesenchymal breast cells maintained long-term in novel ex vivo culture conditions in serum free medium. 2.) To analyze changes in gene expression profiles of normal human primary epithelial and stromal/mesenchymal breast cells upon long-term ex vivo co-culture when compared to corresponding monocultures 3.) To study the dynamic reciprocity between normal human primary epithelial and stromal/mesenchymal breast cells. 4.) To identify critical molecular pathways and biomarkers controlling epithelial and/or stromal cell growth and quiescence. Human primary epithelial progenitor cells and mesenchymal stem cells bearing fluorescent tags were either co-cultured in novel ex vivo culture conditions on ECM coated meshes in serum free medium (M5) or cultured as monocultures in the same conditions for 30 days. The cultures were then dissociated and epithelial and stromal/mesenchymal cells from either co-cultures or monocultures separated by FACS. Gene expression profiling of epithelial or stromal/mesenchymal cells was performed. Clean gene expression profiles from three different epithelial and stromal/mesenchymal cell extracts either grown in co-cultures or monocultures were successfully obtained.

2015-09-26 | GSE51201 | GEO

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