The determination of cell surface phenotypes of lps-stimulated u937-derived microparticles
Microparticles (MP) are cell vesicles that are produced by cells such as endothelial cells and platelets. These MP productions are triggered under certain circumstances such as apoptosis and activation of cells. The purpose of this project was to characterize the cell surface phenotype of monocyt...
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| Format: | Monograph |
| Language: | English |
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Universiti Sains Malaysia
2016
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| Online Access: | http://eprints.usm.my/62259/ http://eprints.usm.my/62259/1/NUR%20HIDAYAH%20BINTI%20JAMALUDIN%20-%20e.pdf |
| Summary: | Microparticles (MP) are cell vesicles that are produced by cells such as endothelial cells
and platelets. These MP productions are triggered under certain circumstances such as
apoptosis and activation of cells. The purpose of this project was to characterize the cell
surface phenotype of monocytic MP (mMP) derived from 1)937 cells after being stimulated
with LPS. In this study, U937 cells were stimulated with 1 ug/ml LPS for 18 and 24 hours.
After LPS stimulation, cell viability was detected using Trypan blue exclusion on LPSstimulated
U937 cells. Both cell pellet and mMP were stained with CD14, CD11b and HLADR
before being analysed by flow cytometry. As for mMP detection, cells were co-labeled
with Annexin-V. Flow cytometry analysis showed that both U937 cells and mMP express
CD11b and HLA-DR with slight expression of CD14. As mMP were originated from U937
cells, it is possible for both cells and mMP to express the same surface phenotypes. Furthermore, mMP were also positive for Annexin-V which is a marker of
phosphatidylserine (PS). This PS is a structure that belongs to the cell surface which is
exposed after cell membrane asymmetry being destroyed in apoptosis by LPS. As mMP
structure was derived from the cell membrane of U937 cells, it is possible for mMP to have
PS expression on their surfaces which resembled their mother cells. In summary, LPS
stimulated U937 cells to produce mMP under the condition of cell death or apoptosis as
LPS mimicking inflammation. In future, these mMP can act as specific biomarkers for
inflammation and this project could be the preliminary studies that characterized the mMP. |
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