0000000000513553
AUTHOR
Līga Saulīte
Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages.
Abstract. Background Macrophages are one of the most important players in the tumor microenvironment. The polarization status of tumor associated macrophages into a pro-inflammatory type M1 or anti-inflammatory type M2 may influence cancer progression and patient survival. Extracellular vesicles (EVs) are membrane-bound vesicles containing different biomolecules that are involved in cell to cell signal transfer. Accumulating evidence suggests that cancer-derived EVs are taken up by macrophages and modulate their phenotype and cytokine profile. However, the interactions of cancer-derived EVs with monocytes and macrophages at various differentiation and polarization states are poorly understo…
Additional file 5: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
Effect of SW480 and SW620-derived EVs on biomolecule secretion patterns of monocytes and M0, M1 and M2 macrophages. Luminex data analysis showing TNFα, IL-6, CXCL10, IL-23, IL-10, MMP9, IL-1β and CCL22 concentration in cell culture supernatants of monocytes (M) and M0, M1 and M2 macrophages following incubation with SW480 and SW620 EVs or without them (control). The graphs represent mean ± SD (n = 3). (PDF 39 kb)
Additional file 3: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
Effect of temperature on the SW480 EV uptake in THP-1 monocytes. Flow cytometry histograms showing Syto RNA Select fluorescence intensities of untreated (left) and Syto RNA Select-labeled SW480 EV-treated THP-1 monocytes following incubation at 4 °C (middle) and 37 °C (right). Histogram markers show the percentage of Syto RNA Select-positive cells. (PDF 53 kb)
Ādas mezenhimālo cilmes šūnu fenotipa raksturojums pēc neirotrofo faktoru inducētas neirodiferenciācijas
Ādas mezenhimālās cilmes šūnas (Ā-MCš), pakļautas specifiskiem faktoriem, spēj diferencēties par neironiem un glijas šūnām. Maģistra darba mērķis bija analizēt Ā-MCš fenotipu pēc neirotrofo faktoru inducētas neirodiferenciācijas. Neiroektodermas gēnu ekspresiju Ā-MCš analizēja ar qPCR, cilmes šūnu pluripotences un mezenhimālo marķieru ekspresiju ar plūsmas citometriju, Sigma-1 receptora ekspresiju (SigmaR1) ar qPCR un IF, kā arī BDNF sekrēciju ar ELISA. Ā-MCš fenotipu salīdzināja ar ReNcell CX šūnām. Retīnskābes indukcijas barotnē novēroja nervu kores gēnu ekspresijas pieaugumu. Cilmes šūnu marķieru ekspresija pēc diferenciācijas kritās. Mezenhimālo marķieru ekspresija samazinājās vienā no …
Neirotrofo faktoru ietekme uz neiroektodermas marķieru ekspresiju ādas mezenhimālajās cilmes šūnās
Kultivēšanas barotnes sastāvs ir faktors, kas stimulē cilmes šūnu diferenciāciju in vitro. Neirotrofie faktori virza diferenciāciju neiroektodermas šūnu virzienā. Bakalaura darba mērķis bija analizēt neirotrofo faktoru ietekmi uz neiroektodermas marķieru ekspresiju ādas mezenhimālajās cilmes šūnās (Ā-MCš). Neiroektodermas gēnu ekspresija Ā-MCš tika noteikta ar RT-PCR metodi pēc diferencēšanas trīs barotnēs. Tika izveidots 15 neiroektodermas gēnu praimeru panelis. Sox10, NES un TUBB3 gēni tika analizēti arī ar kvantitatīvo PCR (qPCR) metodi. RT-PCR rezultāti uzrādīja atšķirības Pax6 un S100β gēnu ekspresijā starp barotņu sastāviem. qPCR analīze parādīja, ka Švāna šūnu barotnē pieauga Sox10 e…
Additional file 2: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
SW480 and SW620-derived EV effect on monocyte (M) and macrophage (M0, M1, M2) viability. a OD values at 450 nm which are in direct proportion of viable cell counts. b SW480 and SW620 EV cytotoxicity on THP-1 monocytes and M0, M1 and M2 macrophages. The graphs represent mean ± SEM (n = 3). Statistical analysis carried out with the t-test. *p ≤ 0.05, **p ≤ 0.01 vs. untreated cell control of the respective monocyte-macrophage cell subset. (PDF 50 kb)
Additional file 4: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
TNFα, IL-23, IL-6, IL-1 β, CXCL10, CCL22, IL-10 and MMP9 secretion profile at different monocyte-macrophage differentiation stages. The graphs represent average biomolecule concentrations SEM (n = 3). Statistical analysis carried out with one-way ANOVA test. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 and **** ≤ 0.0001 vs. untreated cell control of the respective monocyte-macrophage cell subset. (PDF 63 kb)
Additional file 1: of Effect of colorectal cancer-derived extracellular vesicles on the immunophenotype and cytokine secretion profile of monocytes and macrophages
Experimental design of the THP-1 monocyte to macrophage differentiation showing the time points for the addition of EVs and stimulatory molecules. Below the experimental design, representative light microscopy images show morphology of THP-1 monocytes (M), M0 macrophages (M0), M1 macrophages (M1) and M2 macrophages (M2) (n = 4). Scale bar 100 μm. Representative flow cytometry dot plots show CD14, HLA-DR, CD206 and CD68 marker expression at M, M0, M1 and M2 stages. (JPG 1845 kb)