CD146+CD45-CD31- pericyte RNA sequencing following an acute bout of electrical stimulation
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ABSTRACT:
PROVIDER: PRJNA505805 | ENA |
REPOSITORIES: ENA
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Project description:The objective of this study was to understand the mechanisms by which electrical stimulation in vivo leads to improvements and growth in skeletal muscle post exercise through changing the function of CD146+CD45-CD31- pericytes. We utilized global gene profiling to identify novel signaling pathways and patterns of gene expression involved in this adaptation process.
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Project description:Human pericytes demonstrate multilineage differentiation potential, and their descendants participate in tissue homeostasis and repair. Increasing evidence from developmental biology and tissue engineering suggest that regional specification by tissue of origin exists among human pericytes. Here, we sought to define the differentiation of CD146+ human pericytes from skeletal and soft tissue sources. Uncultured CD146+CD31-CD45- pericytes were derived by fluorescent activated cell sorting from human periosteum, adipose, or dermal tissue. Periosteal CD146+CD31-CD45- cells retained canonical features of pericytes, including cell surface marker expression, multilineage differentiation potential, and paracrine induced tubulogenesis. Periosteal pericytes demonstrated a striking tendency to undergo osteoblastogenesis, while soft tissue pericytes did not in vitro or in vivo. Microarray analysis demonstrated substantive differences between periosteal pericytes in comparison to their soft tissue pericyte counterparts. In sum, skeletal and soft tissue pericytes differ in their relative lineage differentiation potential and ability to form bone. Human tissues were microdissected, digested, and FACS sorted to derived a CD146+CD31-CD45- pericyte population, and cells were expanded in standard growth medium (DMEM, 10% FBS, 1% pen/strep) and nucleic acid isolated at subconfluency
2021-08-01 | GSE118962 | GEO
Project description:Human pericytes demonstrate multilineage differentiation potential, and their descendants participate in tissue homeostasis and repair. Increasing evidence from developmental biology and tissue engineering suggest that regional specification by tissue of origin exists among human pericytes.Here, we sought to define the differentiation of CD146+ human pericytes from skeletal and soft tissue sources. Uncultured CD146+CD31-CD45- pericytes were derived by fluorescent activated cell sorting from human periosteum, adipose, or dermal tissue. Periosteal CD146+CD31-CD45- cells retained canonical features of pericytes, including cell surface marker expression, multilineage differentiation potential, and paracrine induced tubulogenesis. Periosteal pericytes demonstrated a striking tendency to undergo osteoblastogenesis, while soft tissue pericytes did not in vitro or in vivo. Microarray analysis demonstrated enrichment in CXCR4 and BMP signaling among periosteal pericytes in comparison to their soft tissue pericyte counterparts. Signaling pathway manipulation of CXCR4 among soft tissue pericytes led to an increase in osteoblastogenesis and bone formation. In sum, skeletal and soft tissue pericytes differ in their relative lineage differentiation potential and ability to form bone. Plasticity exists, however, and manipulation of CXCR4 signaling pathway may ‘coax’ a soft tissue pericyte toward an osteoblastogenic cell fate.
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Project description:Single cell RNA sequencing for bone marrow CD31+CD45- cells from diaphysis, metaphysis or epiphysis of mouse femurs
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Project description:We report the RNAseq data obtained from 50.000-100.000 CD31-/CD45- pneumocytes isolated by FACS from mice harboring a normal dose or one extra copy of the Sirt1 gene, and a tamoxifen-inducible oncogenic KI alelle of KRasG12V after 4 weeks of tamoxifen treatment. Pneumocytes with the activated form of the inducible KRasG12V oncogene sere selected making use of the reporter gene LacZ (located next to the oncogene in the same polycistronic mRNA), by loading CD31-/CD45- pneumocytes with the LacZ-activated fuorogenic molecule FDG prior to FACS sorting.
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Project description:To investigate the heterogeneity of lung stromal cells and identify the specific lung stromal subset, we performed single cell RNA-sequencing (scRNA-seq) on lung stromal cells (CD45-CD31-CD326-). Around 6800 cells were captured using the 10x Chromium technology.
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Project description:To understand if cancer cells that metastasize to the lung induce changes of gene expression in associated endothelial cells to promote metastatic growth, we profiled the gene expression of CD31+/CD45- cells sorted from unchallenged normal mouse lungs and dissected metastatic nodules.
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2021-08-24 | PXD026963 | Pride
Project description:A major challenge in realizing regenerative treatment using freshly isolated Adipose Derived Regenerative Cells (ADRCs) is the cellular heterogeneity and donor variability. Ex vivo expanded ADRCs (=ASCs) are available at larger quantities and represent a more homogeneous population suitable for allogenic use. Emerging pre-clinical and clinical data however suggest similar, or even superior efficacy of ADRCs as compared to ASCs for indications involving (micro)vascular deficiency. Since CD31+ ADRCs lack in cultured ASCs, we hypothesized that these cells account for improvement of vascular function in the heterogenous ADRCs. Herein, we found that in patients with erectile dysfunction (ED), the number of injected CD31+ ADRCs correlates positively with erectile function 12 months after one bolus of autologous ADRCs. Comprehensive in vitro and ex vivo analyses confirmed superior pro-angiogenic and paracrine effects of human CD31+ enriched ADRCs compared to the corresponding CD31- and parent ADRCs. CD31+, CD31- and ADRCs were co-cultured in aortic ring-, as well as in ED-relevant corpus cavernousum tube formation assays, both showing a significantly higher ability of the CD31+ ADRCs to support tube development. This effect was corroborated using conditioned medium (CM), while quantitative mass spectrometric analysis suggested that this is explained by secretory pro-angiogenic proteins such as DKK3, ANGPT2, ANAX2 and VIM, all being enriched in CD31+ADRC CM. To gain further specification of the ADRC subpopulations expressing these proteins, single-cell RNA sequencing was performed and showed that transcripts of the upregulated and secreted proteins were present in 9 endothelial ADRC subsets including endothelial progenitor cells in the heterogenous non-cultured ADRCs. Our data thus suggest that the vascular benefit of using ADRCs in regenerative medicine is dictated by CD31+ ADRCs, which likely represent an improved alternative to heterogenous ADRCs as well as ASCs when aiming for vascular repair in cell therapy.
2023-10-24 | PXD040388 | Pride