Project description:Next-generation sequencing (RNAseq) analysis of gene expression changes during the long-term in vitro culture and osteogenic and chondrogenic differentiation of ADSCs remains to be important, as the analysis provides important clues toward employing stem cells as a therapeutic intervention. In this study, the cells were isolated from adipose tissue obtained during routine surgical procedures and subjected to 14-day in vitro culture and differentiation. The mRNA transcript levels were evaluated using the Illumina platform, resulting in the detection of 19,856 gene transcripts. The most differentially expressed genes (fold change >|2|, adjusted p value < 0.05), between day 1, day 14 and differentiated cell cultures were extracted and subjected to bioinformatical analysis based on the R programming language. The results of this study provide molecular insight into the processes that occur during long-term in vitro culture, osteogenic and chondrogenic differentiation of ADSCs, allowing the re-evaluation of the roles of some genes in MSC progression towards a range of lineages. The results improve the knowledge of the molecular mechanisms associated with long-term in vitro culture and differentiation of ADSCs, as well as providing a point of reference for potential in vivo and clinical studies regarding these cells' application in regenerative medicine.
Project description:We analyzed the difference of gene expression between the isolates havested from long-term cultures (Ma et.al. 2020). In this study, the paradaux cell line were cultured for over 40 days under different population control conditions (uncontrolled, negative feedback and paradoxical feedback. Isolates of each culture were harvested at the end of the long-term culture and preped for whole genomic RNA sequencing.
Project description:Transcription profiling by high throughput sequencing of human polarized cerebellar plate neuroepithelium in three-dimensional culture
Project description:Extracellular vesicles isolated from the conditioned media of short, long-term, and rapamycin treated long-term cultured astrocytes were subjected to proteomic analysis to determine the how the EV proteome changes following long-term culture.
2020-05-27 | PXD017058 | Pride
Project description:Whole genome re-sequencing for diatom under long-term ocean warming
Project description:Expression analysis of human melanoma short-term culture WM451-Lu harvested after lentiviral infection with a GFP (control) or SETDB1 (experimental) viral vector
Project description:Naïve mouse ESCs exhibit full term developmental competence thus hold great potential in regenerative medicine. Maintaining genome stability is essential for potential application of ESCs in stem cell therapy. While ESC potency fluctuate with retrovirus activity, it is unclear whether long-term cultures affect retrotransposition and genomic stability. We compared retrotransposons in naïve ESCs using various approaches. We show that feeder-serum based culture conditions and small molecule LCDM maintain appropriate expression of retrovirus activity following long-term cultures, whereas cultures under 2iLif and a2i result in aberrant upregulation of retrotransposons. This leads to high frequent retrotransposition. Moreover, naïve ESCs on feeder-serum based culture conditions and small molecule LCDM still generate complete ESC pups by TEC assay, functional test of pluripotency, following long-term cultures, despite that all culture conditions maintain high expression levels of pluripotent genes such as Oct4, Nanog, Klf4 and Sox2. Meanwhile, long telomeres are maintained in ESCs under cultures in Feeder-serum and LCDM conditions but telomeres shortened in other conditions with increasing passages. These data provide insights into retrotransposition, genomic stability and pluripotency of naïve ESCs.