Project description:To date, different experimental strategies have been developed for the ex vivo expansion of human hematopoietic stem (HSCs) in clinical application. However, it is still unclear to what difference in genomic function in HSCs expansion under different culture systems. In this study, we compared gene-expression profile of ex vivo expanded serum (10% FBS, fetal bovine serum) and serum-free culture systems, and then analyze molecular function of differentially expressed gene using microarray chips.
2021-09-01 | GSE126909 | GEO
Project description:Impact of long-term fertilization on fungal communities in one at the long term static fertilization experiment in Bad Lauchstadt
Project description:The value of synthetic microbial communities in biotechnology is gaining traction due to their ability to undertake more complex metabolic tasks than monocultures. However, a thorough understanding of strain interactions, productivity and stability is often required to optimize growth and scale up cultivation. Quantitative proteomics can provide valuable insights into how microbial strains adapt to changing conditions in biomanufacturing or bioremediation scenarios. However, current workflows and methodologies are not suitable for simple artificial co-culture systems where strain ratios are dynamic. Here, we established a standard workflow for co-culture proteomics using an exemplar system containing two key members, Azotobacter vinelandii and Synechococcus elongatus. Factors affecting the quantitative accuracy of co-culture proteomics were investigated, including peptide physicochemical characteristics such as molecular weight, isoelectric point, hydrophobicity, and dynamic range, as well as factors relating to protein identification such as varying proteome size and shared peptides between species. Different quantification methods based on spectral counts and intensity were evaluated, demonstrating good correlations between protein amount and the six quantification methods at the protein level. We propose a new normalization method, named “LFQRatio”, to reflect the relative contributions of the two distinct cell types emerging from the cell ratio changes during co-cultivation. LFQRatio can be applied to real co-culture proteomics experiments, providing accurate insights into quantitative proteome changes in each strain.
2023-11-02 | PXD046627 | iProX
Project description:Soil prokaryotic and fungal communities in long-term N fertilization trial
| PRJNA736632 | ENA
Project description:Study of fungal community in paddy soil under long-term fertilization
Project description:mRNA sequencing of mesenchymal stem cells in 2D culture systems, mesenchymal stem cells spheroids and mesenchymal stem cells/extracellular matrix in 3D culture systems to profile gene expressions
Project description:Organoids retain the morphological and molecular patterns of their tissue of origin, are self-organizing, relatively simple to handle and accessible to genetic engineering. Thus, they represent an optimal tool for studying mechanisms of tissue maintenance and aging. Long-term expansion under standard growth conditions, however, is accompanied by changes in growth pattern and kinetics. As a potential explanation of these alterations, epigenetic drifts in organoid culture have been suggested. Here, we study histone tri-methylation at lysine 4 (H3K4me3) and 27 (H3K27me3) and transcriptome profiles of intestinal organoids derived from mismatch repair (MMR)-deficient and control mice and cultured for 3 and 20 weeks, and compare them with data on their tissue of origin. We find that, besides the expected changes in short-term culture, organoids show profound changes in their epigenome also during long-term culture. The most prominent are epigenetic gene activation by H3K4me3 recruitment to previously unmodified genes and by H3K27me3 loss from originally bivalent genes. We show that long-term culture is linked to broad transcriptional changes that indicate an ongoing maturation and metabolic adaptation process. This process is disturbed in MMR-deficient mice, resulting in endoplasmic reticulum (ER)-stress and Wnt-activation. Our results can be explained in terms of a mathematical model assuming that epigenetic changes during long-term culture involve DNA de-methylation that ceases if the metabolic adaptation is disturbed.
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.