Primary cells were isolated from 686LN xenografted tumors. After the dissection of tumors, single cells were isolated using tumor tissue dissociation kit (Miltenyi Biotech) then initially positively sorted using CD11b microbeads using a MACS seperator and MS columns (Miltenyi Biotech). CD11b+ population were further sorted using a FACS Aria cells were isolated using magnetic beads (Miltenyi Biotech). Macrophages were sorted using FACS Aria (BD Biosciences). Cells were labeled with fluorochrome-conjugated antibodies, PE/Cy7 anti-mouse Ly-6G, PerCP/Cy5.5 anti-mouse CD11b (Biolegend).
Tumor tissue dissociation kit
Manufactured by Miltenyi Biotec
Sourced in Germany
The Tumor Tissue Dissociation Kit is a laboratory product designed to facilitate the dissociation of tumor tissue samples into single-cell suspensions. The kit contains reagents and protocols to enable the efficient extraction of cells from solid tumor samples for further analysis or downstream applications.
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Lab products found in correlation
Pe cy7 anti mouse ly 6g, by BioLegend (1 mentions)
Macs seperator, by Miltenyi Biotec (1 mentions)
Percp cy5.5 anti mouse cd11b, by BioLegend (1 mentions)
Facs aria, by Miltenyi Biotec (1 mentions)
Ms column, by Miltenyi Biotec (1 mentions)
Magnetic bead, by Miltenyi Biotec (1 mentions)
Facsaria, by BD (1 mentions)
Gentlemacs tissue processor, by Miltenyi Biotec (1 mentions)
Anti cd326 microbeads, by Miltenyi Biotec (1 mentions)
Epcam, by BioLegend (1 mentions)
2 protocols using tumor tissue dissociation kit
1
Isolation of Tumor-Associated Macrophages
2
Isolation and Characterization of CRC Cells
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Primary tumor tissues and metastatic LN tissues were aseptically harvested from CRCLNM patients or HCT116 CRCLNM xenografts, cut into small pieces (∼1 mm3 in size), and digested with the tumor tissue dissociation kit (Miltenyi Biotec, Kolner, Germany, 130-108-339). Single-cell suspension of digested tissues was obtained using a GentleMACS™ tissue processor (Miltenyi Biotec). Primary CRC (P-CRC) cells, LNM-CRC cells, HCT116P and HCT116LNM cells were filtered with a 100 μm filter, collected with anti-CD326 microbeads (Miltenyi Biotec., 130-061-101), and characterized as positive for EpCAM (Biolegend, CA, USA, 369809) and CK19 (R&D, MN, USA, AF3506) with purity >95%, as determined by flow cytometry.
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