Ascites Single Cell Processing

After sample extraction (Figures 1C, D), the fresh cells samples were immediately stored in the sCelLiVE® Tissue Preservation Solution (Singleron) on ice. The sample of ascites were transferred to a 15-ml centrifuge tube. The samples were then filtered with 40 µm sterile strainers, and centrifuged at 1,000 rpm at 4°C for 5 min. Next, 2 ml GEXSCOPE® red blood cell lysis buffer (RCLB, Singleron) was added to lyse the red blood cells for 10 min. Finally, the single cell suspension was collected after re-suspension with PBS, and trypan blue (Sigma) staining was used to calculate cell activity and cell count under a microscope (7 (link)).

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Yu B., Jing P., Gao F., Zhang P., Zheng G, & Zhang X. (2024). Effect of sodium stibogluconate in recruiting and awakening immune cells in the pleural fluid of pancreatic cancer: preparation for immunotherapy. Frontiers in Immunology, 14, 1315468.

Publication 2023

sCelLiVE® Tissue Preservation Solution GEXSCOPE® red blood cell lysis buffer (RCLB, trypan blue

Corresponding Organization : Peking University Cancer Hospital

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Variable analysis

independent variables

Sample extraction method (using sCelLiVE® Tissue Preservation Solution)
Filtration of samples with 40 µm sterile strainers
Centrifugation of samples at 1,000 rpm at 4°C for 5 min
Red blood cell lysis using GEXSCOPE® red blood cell lysis buffer (RCLB) for 10 min

dependent variables

Cell activity
Cell count

control variables

Temperature (4°C)
Centrifugation speed (1,000 rpm)
Centrifugation duration (5 min)
Red blood cell lysis duration (10 min)

controls

None specified
None specified

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