Project description:Primary cells deficient for PDCD10/CCM3 do not enter senescence as control cells. Microarray analysis was performed in cells transduced with non-targeting shRNA and CCM3 shRNA at passage 7 (early passage) and passage 11 (late passage), when control cells are already senescent.
Project description:A MIBC patient with aggressive disease course was selected for ex vivo model establishment and growth dynamic studies. After standard neoadjuvant treatment with gemcitabine and cisplatin, cystectomy was performed, and excised urothelial carcinoma tissue was implanted subcutaneously into male SCID mice to generate CoCaB1 PDXs. Established PDXs were passaged subcutaneously in SCID mice to generate early and late passage models for functional and molecular characterization. Representative early and late passage PDXs were collected for organoid establishment.
Project description:Primary cells deficient for PDCD10/CCM3 do not enter senescence as control cells. Microarray analysis was performed in cells transduced with non-targeting shRNA and CCM3 shRNA at passage 7 (early passage) and passage 11 (late passage), when control cells are already senescent. Primary endothelial cells were transduced either with non-target shRNA or with CCM3 shRNA. RNA was extracted at passage 7 and passage 11
Project description:We performed microarray analysis to detect the differential expression of miRNAs in early- vs. late-passage DP cells by microarray profiling of the total of 1924 hsa-miRNAs (miRNAs)
Project description:Transcription profiling of human mesenchymal cells (MSCs) after ex vivo expansion under culture conditions supporting extensive cell proliferation. Highly efficient cell expansion within short time (~40 days) and comparison of gene expression profiles from MSCs after primary seeding, defined as early passage versus MSCs after a minimum of 17 (max 34) population doublings, defined as late passage. Combined data of passage 1 and 2 was compared to passage 0 (zero) as reference
Project description:Bone marrow stromal cells (BMSCs) can be expanded by serial passage, but expansion is limited by cell senescence. The nature of changes associated with BMSC serial passages was assessed. Transcriptome analysis of 10 early and 15 late passage samples from 5 subjects revealed 2193 differentially expressed genes; those highly expressed in early passage cells were overrepresented in skeletal system development, embryonic morphogenesis, tube morphogenesis, etc, while those highly expressed in the late passage BMSCs were overrepresented in nucleosome assembly; chromatin assembly, DNA packaging, etc. 57 BMSC samples from 7 donors were further analyzed for the transition from an early to late passage; 155 genes were highly correlated with BMSC senescence and a set of 24 genes was predictive of BMSCs lifespan. The change from an early to a late passage molecular signature occurred between passage 3 and 5. In contrast, senescence associated beta-galactosidase staining began to increase after passage 6 or 7 and colony formation efficiency began to fall after passage 7. These data indicated that the onset of molecular changes associated with BMSC passage varied among individuals and preceded changes in commonly used indicators of BMSC senescence. The set of 24 BMSC lifespan predictive genes will be useful in assessing the quality of clinical BMSC products.
