Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA116511 | ENA |
REPOSITORIES: ENA
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Project description:Quantitation of polyA mRNA levels in subpopulations of the HexVenus reporter (clone HV5.1) mouse ES cell line growing under self-renewing conditions. Subpopulations were identified and isolated based on the expression of the ES cell surface marker SSEA 1 and the expression of the venus protein. At approximately 70% confluence, cells were trypsinised, resuspended in FACs buffer (10%FCS in PBS) and incubated with a mouse monoclonal antibody to SSEA 1 (MC-480, Developmental Hybridoma Studies Bank, University of Iowa). Cells were then incubated with an Alexa-647 conjugated anti-mouse IgM antibody (Invitrogen) and subpopulations were fractionated by flow cytometry. The aim was to identify genes which are differentially expressed between FACS-sorted HV-S+ and HV+S+ primed mESC populations.
2016-10-11 | GSE64939 | GEO
Project description:Most differentiation protocols for generation of hepatocyte-like cells from iPS cells generate cells with heterogenous expression of hepatic markers, which confounds results from liver disease models involving complex traits and subtle phenotypes We utilized proteomics to identify heptocyte-restricted cell surface proteins that mark a subpopulation of iPS cell derived hepatocytes. By performing microarray on FACS sorted cells, we demonstrate that subpopulation of hepatocyte-like cells expressing SLC10A1 are enriched for hepatic markers
2016-12-22 | GSE80279 | GEO
Project description:In the context of most induced pluripotent stem (iPS) cell reprogramming methods, heterogeneous populations of nonproductive and staggered productive intermediates arise at different reprogramming time points1-11. Despite recent reports claiming substantially increased reprogramming efficiencies using genetically modified donor cells12,13 prospectively isolating distinct reprogramming intermediates remains an important goal to decipher reprogramming mechanisms. Previous attempts to identify surface markers of intermediate cell populations were based on the assumption that during reprogramming the cells progressively lose donor cell identity and gradually acquire iPS cell properties1,2,7,8,10. Here, we report that iPS cell and epithelial markers, such as SSEA1 and EpCAM, respectively, are not predictive of reprogramming during early phases. Instead, in a systematic functional surface marker screen we find that early reprogramming-prone cells express a unique set of surface markers, including CD73, CD49d and CD200 that are absent in fibroblasts and iPS cells. Single cell mass cytometry and prospective isolation show that these distinct intermediates are transient and bridge the gap between donor cell silencing and pluripotency marker acquisition during the early, presumably stochastic reprogramming phase2. Expression profiling revealed that the transcriptional regulators Nr0b1 and Etv5 are specifically expressed in this early reprogramming state, preceding activation of key pluripotency regulators such as Rex1, Dppa2, Nanog and Sox2. Both factors are required for the generation of the early intermediate state and fully reprogrammed iPS cells, and thus mark some of the earliest known regulators of iPS cell induction. Our study shows an ordered sequence of transitions during the earliest steps of iPS cell reprogramming that deconvolutes the first steps in a hierarchical series of events that lead to pluripotency acquisition. Samples for poised (CD73+ or CD49d+) and non-poised (CD73-) reprogramming samples were FACS sorted 6 and 9 days after induction of Klf4, Oct4, Sox2 and cMyc in Rosa-rtTA +/- mouse embryonic fibroblasts (MEFs). 'Total' populations are expression analyses for unsorted populations analyzed at the same time points. Control populations were also sampled: mouse embryonic fibroblasts (MEFs), partially reprogrammed cells (SC4) and mouse embryonic stem cell (ESC).
2015-01-31 | E-GEOD-62957 | biostudies-arrayexpress
Project description:Transcriptomic profiling of second heart field subpopulations FACS sorted
2020-08-31 | GSE123771 | GEO
Project description:Fluorescence-activated cell sorting (FACS) is a specialized technique to isolate
cell subpopulations with a high level of recovery and accuracy. However, the cell sorting
procedure can impact the viability and metabolic state of cells. Here, we performed a comparative study and evaluated the impact of traditional high-pressure charged droplet-based and a microfluidic chip-based sorting approach on the metabolic and phosphoproteomic profile of different cell types. While microfluidic chip-based sorted cells more closely resembled the unsorted control group for most cell types tested, the droplet-based sorted cells showed significant metabolic and phosphoproteomic alterations. In particular, greater changes in redox and energy status were present in cells sorted with the droplet-based cell sorter along with higher transcriptional and spliceosomal regulation and mechanical stress signaling. These results
indicate microfluidic chip-based sorting is less disruptive compared to droplet-based sorting.
2023-11-09 | MSV000093348 | MassIVE
Project description:Profiling of open chromatin on second heart field subpopulations FACS sorted
2020-08-18 | GSE123765 | GEO
Project description:By combination of FACS sorting based on lipid sensitive Nile Red staining and small cell number proteomics the occurrence of population heterogeneity was analyzed. First population heterogeneity was identified by Nile Red staining and FACS in N. oceanica cells grown in nitrogen replete and nitrogen deplete media. For each condition two subpopulations(+NP3,+NP4,-NP3,-NP4) were detected and three replicates were sorted. Using the gathered proteome data proteins were identified and quantified with the MaxQuant software. Based on these quantification results a two sample t-test was performed to show significant regulation between +NP3 vs. +NP4 or -NP3 vs. -NP4. Goal was to understand the biological reasons for differences in lipid production between subpopulations.
2019-02-15 | PXD008721 | Pride
Project description:Organoids derived from a human small intestine were altered so that cells producing the GIP peptide would generate a flouresence marker. These cells were then sorted based on the fluorescence in a FACS system and positive cells collected along wiht negative cell population. These cells were then lysed and analysed using a peptidomics approach
2024-11-15 | PXD052659 | Pride
Project description:B cell plays an important role in antibody induction response. We were aiming to describe two distinct B cell subpopulations in the intestinal lymphoid tissue ‘Peyer’s patches’ of mice, characterized by their transcriptional profile. B cells were sorted according to a combination of surface marker from wild type young mice, that were treated with bacteria for 7 days or control. RNA was extracted. Microarray analysis was then applied to identify the difference between two B cell subsets and the changes of expression within each B subset, due to treatment.
2018-12-31 | GSE114571 | GEO
Project description:The cellular heterogeneity of one patient derived orthotopic breast cancer xenograft model (PDBCX) was investigated using flow cytometry , combined with assessment of in vivo tumorigenicity and whole genome expression profiling. Epithelial cell adhesion molecule (EpCAM) was revealed as a highly specific cell surface marker of the human tumor cell population in both xenografts. Based on expression patterns observed in primary tumor tissue, SSEA-4 and CD24 were chosen as markers to further subdivide the luminal tumor cells into four subpopulations. FACS sorting was used to isolate four cell subpopulations. Results: In vivo tumorigenicity assay showed that SSEA-4+/CD24+ cells were non-tumorigenic, while the three other subpopulations were tumorigenic. Tumors resulting from the SSEA-4+/CD24- subpopulation of luminal cancer cells, did not express CD24, while tumors arising from the SSEA-4-/CD24-, and SSEA-4-/CD24+ populations both recapitulated the original tumor containing all four subpopulations. Whole genome expression analysis revealed distinct transcriptional profiles, and 44 genes were significantly differentially expressed when comparing the tumorigenic vs non-tumorigenic populations. Several interesting genes putatively suppressing the cancer cells ability to initiate tumors in vivo were upregulated in the non-tumorigenic population. We here show that tumor initiating cells within one primary tumor evidently included more than one phenotype. Furthermore, with respect to cell surface marker expression, one of the subpopulations produced tumors unlike both the originating cells, and the original tumor. Discussion: Our results imply that subpopulations from one primary tumor can give rise to dissimilar daughter tumors. These tumors may not necessarily respond to the same targeted treatment, and thereby represent a therapy escape mechanism. This study highlights that to remove the risk of breast cancer recurrence, inhibition of the molecules critical for driving the tumor progression in several tumor cell subpopulations might be essential Gene expression was measured in four cell subpopulations isolated from Patient derived human luminal-like breast cancer xenograft. Four replicates from three subpopulations and three replicates from one subpopulation.
2015-05-26 | E-GEOD-48384 | biostudies-arrayexpress