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課題案例系列之：
外泌體（Exosomes）
案例簡介>>
外泌體（exosomes）是直徑在100nm左右，來源于內體（endosome）細胞質膜內陷形成的多泡體，多泡體與其他細胞內囊泡和細胞器融合，為外泌體的形成提供了不同類型的原料。
外泌體作為細胞間通訊的工具， 在體液中游走， 到達靶點后， 可以 作用于另一種細胞， 引發(fā)受體細胞產生應答。外泌體對受體細胞作用目 前已有很多相關研究。細胞吸收和分泌外泌體的過程彼此相聯(lián)系，外泌 體不同的吸收途徑和機制以及對不同類型細胞作用的特異性增加了外泌 體在細胞間通訊功能的復雜性。外泌體在發(fā)育， 免疫應答，疾病發(fā)生， 組織器官修復等生物學過程中均發(fā)揮著重要作用。
技術路線>>
結果展示>> 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;
技術路線>>
結果展示>> 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.
???技術路線>>
結果展示>> 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;
技術路線>>
結果展示>> 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.