Project description:To investigate the transcriptional regulation by p53 in astrocytes, we performed ChIP-sequencing using p53 antibody (Leica CM-5) or rabbit IgG control in wildtype primary mouse cortical astrocytes.
Project description:To investigate the mechansims that underly astrocyte dedifferentiation, we performed single cell RNA sequencing analysis of primary astrocytes after p53 loss and exposure to mitogens, EGF and and FGF. Primary astrocytes were isolated from postnatal day 3 inducible p53 knockout mice (GFAP-CreERT2;p53flox/flox;LSL-tdTomato), whereby treatment with 4-hydroxytamoxifen (4OHT) induces p53 loss and tdTomato labelling in GFAP+ astrocytes. Astrocytes were treated with 4OHT in media supplemented with EGF and FGF to induce recombination and astrocyte dedifferentiation in vitro.
Project description:To investigate the mechansims that underlie astrocyte dedifferentiation, we performed single cell RNA sequencing analysis of primary astrocytes isolated from adult mouse cortex after p53 loss and exposure to mitogens, EGF and and FGF. Primary astrocytes were isolated from cortices of 2-3month old inducible p53 knockout mice (GFAP-CreERT2;p53flox/flox;LSL-tdTomato) using Miltenyi Adult Brain Dissociation kit and ACSA-2 beads. Treatment with 4-hydroxytamoxifen (4OHT) induces p53 loss and tdTomato labelling in GFAP+ astrocytes. Astrocytes were treated with 4OHT in media supplemented with EGF and FGF to induce recombination and astrocyte dedifferentiation in vitro.
Project description:The individualized treatment of tumors has always been an urgent problem in clinical practice. Organoids-on-a-chip can reflect the heterogeneity of tumors and is a good model for in vitro anticancer drug screening. In this study, surgical specimens of patients with advanced colorectal cancer will be collected for organoid culture and organoids-on-a- chip. Use organoids-on-a-chip to screen tumor chemotherapy drugs, compare the results of patients’ actual medication regimens, and study the guiding role of organoids in the formulation of precise tumor treatment plans. The investigators will compare the response of organoids to drugs in vitro with the patient’s response to actual chemotherapy and targeted drugs and explore the feasibility and accuracy of organoids-on-a-chip based drug screening for advanced colorectal cancer. The project will establish a screening platform for chemotherapeutic drugs and targeted drugs based on colorectal cancer organoids to quickly and accurately formulate personalized treatment plans for clinical patients.
Project description:ChIP-seq analysis of Primary cortical astrocytes harboring a single floxed Trp53 allele harvested from male and female postnatal day one pups, and transduced with mutant p53;IRES GFP followed by CRE recombinase. p53 protein was immunoprecipated from fixed whole chromatin from cells expressing one of three mutants or p53 KO control.
Project description:This DNA methylation dataset describes epigenomic changes in in vitro serially passaged primary and immortalized astrocytes, in the context of studies examining cellular aging patterns that are conserved in vivo and in vitro. Primary and fetal hTERT-immortalized astrocytes were grown under normoxic conditions and serially passaged. Longitudinal DNA samples were collected throughout passaging and DNA methylation was measured using the Infinium HumanMethylation850 BeadChip.
Project description:We used microarrays to compare the global programme of gene expression in primary cultures of neurons and astrocytes. These data sets were compared to the expression profiles of other tissues, including pancreatic islets, in order to identify a specific neuroendocrine program in pancreatic islets. Neurons and astrocytes were isolated from brain cortex and cultured in vitro for 7 and 20 days respectively. RNA was extracted and hybridized on Affymetrix microarrays. Two biological samples from neurons and astrocytes were analyzed.