Age dependence of hematopoietic progenitor survival and chemokine family gene induction after gamma-irradiation in bone marrow tissue in C3H mice
ABSTRACT: To identify the age dependent radiation response in C3H mouse bone marrow cells. Changes in gene expression in 1 week old and 8 weeks old C3H mouse (female) bone marrow after 2Gy irradiation. Bone marrow samples were collected un-irradiated control, 6 hours and 24 hours after irradiation. Three mice were used at each time course.
Project description:Human bone marrow mesenchymal stem cells (BMMSC) and human embryonic mesenchymal stem cells (ESMSC)were used. X-ray irradiation (2Gy) or 0Gy was delivered. After 4 hours, equal amount total RNA from each sample was extracted prior to gene expression analysis. After making cDNA, genes associated with Wnt signaling pathway were analyzed. qPCR gene expression profiling. Human bone marrow mesenchymal stem cells and human embryonic mesenchymal stem cells were used. X-ray irradiation (2Gy) or 0Gy was delivered. After 4 hours, equal amount total RNA from each sample was extracted prior to gene expression analysis.
Project description:To elucidate the mechanism of the in vivo radioprotection activity of Zn-containing heat-treated Saccharomyces cerevisiae yeast (Zn-yeast), expression changes in bone marrow after whole body irradiation (WBI) with concomitant treatment of Zn-yeast were analyzed using microarray technology. Keywords: mouse, bone marrow, Zn-containing heat-treated yeast administration, gamma-irradiation Zn-yeast suspension was administered i.p. into C3H mice immediately after 7.5 Gy WBI. Bone marrow from irradiated mice was extracted at 6 hours after irradiation and analyzed by microarray containing of 44,000 probes.
Project description:To elucidate the mechanism of the in vivo radioprotection activity of Zn-containing heat-treated Saccharomyces cerevisiae yeast (Zn-yeast), expression changes in bone marrow after whole body irradiation (WBI) with concomitant treatment of Zn-yeast were analyzed using microarray technology. Keywords: mouse, bone marrow, Zn-containing heat-treated yeast administration, gamma-irradiation Overall design: Zn-yeast suspension was administered i.p. into C3H mice immediately after 7.5 Gy WBI. Bone marrow from irradiated mice was extracted at 6 hours after irradiation and analyzed by microarray containing of 44,000 probes.
Project description:We performed a comprehensive analysis of gene expression changes following low-intensity pulsed ultrasound (LIPUS) treatment of cultured bone marrow cells under bone formation conditions, using cDNA microarray analysis Bone marrow cells were obtained from the femora of rats and were suspended in an osteogenic medium to make a cell culture. After cultures were established, test cultures were exposed to LIPUS via the base of the cell culture plates for 15 min/day on days 3–9 (LIPUS group). Control cultures (without LIPUS exposure) were otherwise treated identically to the LIPUS group. On day 10, total RNA was extracted from both sets of cultures and hybridized to microarray slides, the data sets were analysed. Markers for differentiated osteoblasts and osteocytes, as well as collagen-related genes, cell adhesion factors were up-regulated in LIPUS group on day 10. Gene expression in response to LIPUS treatment of bone marrow cells were measured on day 10 after cultures were established. Experiments were performed at each conditions (LIPUS or Control).
Project description:Mast cells are indispensable for LPS-induced septic hypothermia, in which TNF-α plays an essential role to initiate sepsis. Tec family non-receptor tyrosine kinases ITK and BTK regulate mast cell-derived TNF-α in response to allergic antigen, but their role in LPS-induced TNF-α production by mast cells and related pathology is unclear. We sought to investigate the role(s) of ITK and BTK in mast cell response in septic condition. We found that the absence of ITK and BTK leads to enhanced TNF-α production by bone marrow-derived mast cells (BMMC). Itk-/-Btk-/- mast cells exhibit hyperactive preformed and LPS-induced TNF-α production, along with enhanced expression of other related genes such as NF-κB targeted genes, compared to WT cells. Bone marrow cells from 8-week old WT, Itk-/-, Btk-/- and Itk-/-Btk-/- (double knockout: DKO) C57Bl/6 mice were cultured in murine Interleukin-3/Stem cell factor (IL-3/SCF) supplemented medium for 5 weeks to derive mast cells. WT, Itk-/-, Btk-/- and DKO bone marrow-derived mast cells (BMMC) were factor starved in medium without IL-3/SCF for 12 hours, followed by treatment with PBS (control) or 100 ng/ml LPS for 1 hour. Triplicates of each group were subjected to mouse whole genome genechip microarray analysis. Replicates were randomized on different chips to avoid systematic error.
Project description:In this study, we used C3H/10T1/2 cells, a well known model of myogenic conversion, to study the effect of Six4 knockdown on the expression of genes during fibroblasts to myocytes conversion induced by ectopic expression of MyoD We established C3H/10T1/2 cell line with stable Six4 knockdown by short hairpin RNA (shSix4) vs a control cell line (shLuc) and converted these cells into myogenic lineage by retroviral transduction of plasmid encoding Flag-MyoD-myc (pBABE-MyoD) or empty plasmid (pBABE). Cells were then induced to differentiate for 24 hours before RNA extraction.
Project description:To understand gene expression signatures between wild-type and SFRP5-overexpressing cells To examine genes regulated by SFRP5 expression, sets of microarrays were conducted with Lin- c-kitHi Sca-1+ cells and Lin- c-kitLo Sca-1Lo IL7Rα+ CLP-enriched cells derived from adult bone marrow of SFRP5-transgenic mice or their WT littermates.
Project description:To understand the mechanism by which IRF8 promotes basophil differentiation Murine bone marrow Flt3+ GPs and Flt3- GPs were isolated by cell sorting from wild-type or Irf8 deficient mice. Two independent experiments were performed.
Project description:Antibody-based therapy for cancer is now one of the most successful and important strategies for treating patients with hematological malignancies. However, the lack of efficient tumor-associated antigens restricts the targeting therapy of myeloid leukemia. Analysis of the gene expression proﬁles of primary bone marrow samples from human acute myeloid leukemia (AML) patients or healthy donors was to identify and expand novel targets for the treatment of myeloid leukemias. we found that epithelial cell adhesion molecule (EpCAM) is overexpressed in patients with AML. we analyzed the gene expression proﬁles of bone marrow mononuclear cells from 2 human acute myeloid leukemia (AML) patients and 2 healthy donors using an oligonucleotide microarray, to identify up-regulated genes in AML samples comparing with healthy tissues.
Project description:Mesenchymal stromal cells (MSCs) are multipotent progenitors supporting bone marrow hematopoiesis. MSC have an efficient DNA damage response (DDR) and are consequently reatively radio-resistant cells. Therefore, MSCs are key to hematopoietic reconstitution following total body irradiation (TBI) and bone marrow transplantation (BMT). The bone marrow niche is hypoxic and via the heterodimeric transcription factor Hypoxia-inducible factor-1 (Hif-1), hypoxia enhances the DDR. Using gene knock-down, we have previously shown that the Hif-1α subunit of Hif is involved in MSC radio-resistance, however its exact mechanism of action remains unknown. In order to dissect the involvement of Hif-1α in the DDR, we have generated using CRISPR/Cas9 technology, a stable MS5 mouse MSC cell line lacking Hif-1 expression. Herein, we show that it is the whole Hif-1 transcription factor, and not only the Hif-1α subunit, that modulates the DDR of mouse MSCs, and that this effect is dependent upon the integrity of the DNA binding domain. We have also characterized the Hif-1α-dependent proteomic changes undergone by hypoxic MS5 cells. These findings have important implications for the modulation of MSC radio-resistance in the context of BMT and cancer.