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課題案例系列之：
外泌體（Exosomes）
案例簡(jiǎn)介>>
外泌體（exosomes）是直徑在100nm左右，來(lái)源于內(nèi)體（endosome）細(xì)胞質(zhì)膜內(nèi)陷形成的多泡體，多泡體與其他細(xì)胞內(nèi)囊泡和細(xì)胞器融合，為外泌體的形成提供了不同類型的原料。
外泌體作為細(xì)胞間通訊的工具， 在體液中游走， 到達(dá)靶點(diǎn)后， 可以 作用于另一種細(xì)胞， 引發(fā)受體細(xì)胞產(chǎn)生應(yīng)答。外泌體對(duì)受體細(xì)胞作用目 前已有很多相關(guān)研究。細(xì)胞吸收和分泌外泌體的過(guò)程彼此相聯(lián)系，外泌 體不同的吸收途徑和機(jī)制以及對(duì)不同類型細(xì)胞作用的特異性增加了外泌 體在細(xì)胞間通訊功能的復(fù)雜性。外泌體在發(fā)育， 免疫應(yīng)答，疾病發(fā)生， 組織器官修復(fù)等生物學(xué)過(guò)程中均發(fā)揮著重要作用。
技術(shù)路線>>
結(jié)果展示>> Fig1
A B
C D
A. The CCK8 experiment verified that after co culturing stem cells with model group cells, the inhibition of cell proliferation in the model group was alleviated;
B. Edu flow cytometry detection verified that after co culture of stem cells with model group cells, the proliferation activity of model group cells was restored;
C. The flow cytometry apoptosis experiment verified that after co culturing stem cells with model group cells, the apoptosis rate of the model group cells was alleviated;
D. WB and ELISA experiments verified that after co culture of stem cells with model group cells, there were changes in cell related apoptosis and inflammatory indicators in the model group;
技術(shù)路線>>
結(jié)果展示>> Fig2
A B
C D E
?A. Transmission electron microscopy experiments were conducted to determine whether the separated extracellular vesicles have a standard bilayer membrane structure;
B. The particle size distribution of extracellular vesicles obtained from NTA experimental detection;
C. WB experiment detects the expression of extracellular vesicle related markers;
D. High throughput miRNA sequencing experiments detect differential miRNAs in samples and perform pathway analysis to screen out the most likely miRNA targets;
E. The exosomes are labeled with PKH26, and the cells are stained with ghost pen cyclic peptide. Exosomes tracing technology shows the phagocytosis of the exosomes.
???技術(shù)路線>>
結(jié)果展示>> Fig3
A B C
?D E
A. The RT-qPCR Assay was used to detect changes in the content of target miRNA before and after cell model and miRNA transfection;
B. The CCK8 experiment verified that after miRNA transfection, the inhibition of cell proliferation in the model group was alleviated;
C. Edu flow cytometry confirmed that after co culturing stem cells with model group cells, the proliferation activity of model group cells was restored;
D. After co culturing stem cells with model group cells, the apoptosis rate of the model group cells was alleviated through flow cytometry apoptosis verification;
E. WB and ELISA experiments were conducted to verify that after co culture of stem cells with model group cells, changes in cell related apoptosis and inflammatory indicators occurred in the model group;
技術(shù)路線>>
結(jié)果展示>> Fig4
A B C
D
A. The CCK8 experiment detected the changes in cell proliferation in the model group when model cells were co cultured with stem cells that could secrete extracellular vesicles and stem cells that could not secrete extracellular vesicles;
B. Edu immunofluorescence assay was used to detect the changes in cell proliferation in the model group when the model cells were co cultured with stem cells that could secrete extracellular vesicles and stem cells that could not secrete extracellular vesicles;
C. TUNEL fluorescence Assay was used to detect the changes in cell proliferation in the model group under co cultivation of stem cells that can secrete extracellular vesicles and stem cells that cannot secrete extracellular vesicles;
D. WB and ELISA were used to detect the changes in cell proliferation in the model group when the model cells were co cultured with stem cells that could secrete extracellular vesicles and stem cells that could not secrete extracellular vesicles, respectively; The above results indicate that the repair factors expressed by stem cells are mainly transmitted in the form of extracellular vesicles for the repair of model cells.