Mouse tumor infiltrating lymphocyte isolation kit

Manufactured by Solarbio

Sourced in China

The Mouse Tumor-infiltrating lymphocyte Isolation Kit is a laboratory product designed to isolate lymphocytes from mouse tumor tissue samples. The kit provides the necessary reagents and protocols to perform this isolation procedure.

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Lab products found in correlation

Apc anti mouse f4 80, by BioLegend (1 mentions) Phycoerythrin anti mouse cd274 antibody, by BioLegend (1 mentions) Collagenase type 4, by Merck Group (1 mentions) Facscelesta, by BD (1 mentions) Cell incubator, by Thermo Fisher Scientific (1 mentions) Collagenase 1, by neoFroxx (1 mentions) Dnase 1, by Tiangen Biotech (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Collagenase type 5, by Merck Group (1 mentions)

4 protocols using mouse tumor infiltrating lymphocyte isolation kit

Tumor-Infiltrating Immune Cell Profiling

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Tumors were digested to single cell suspension with Type IV collagenase (Sigma-Aldrich). Mouse Tumor-infiltrating lymphocyte Isolation Kit (Solarbio) was used to remove cancer cells and enrich leukocytes. Peridinin-chlorophyll-protein anti-mouse CD45 antibody (Biolegend) was used to mark the leukocytes. APC anti-mouse F4/80 (Biolegend) and fluorescein isothiocyanate anti-mouse/human CD11b (Biolegend) antibodies were used to mark macrophages. Phycoerythrin anti-mouse CD274 antibody (Biolegend) was used to detect the expression of PD-L1. Stained cells were analyzed by BD FACSCelesta (BD Biosciences). Data were processed by FlowJo.

Zhang R., Yang Y., Dong W., Lin M., He J., Zhang X., Tian T., Yang Y., Chen K., Lei Q.Y., Zhang S., Xu Y, & Lv L. (2022). D-mannose facilitates immunotherapy and radiotherapy of triple-negative breast cancer via degradation of PD-L1. Proceedings of the National Academy of Sciences of the United States of America, 119(8), e2114851119.

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Isolation and Characterization of Tumor-Infiltrating Lymphocytes

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After the tumor-bearing mice were sacrificed, subcutaneous tumors were isolated completely. Weigh 0.5 g of tumor tissues were cut and then added to 3 mL of prewarmed (37 °C) 1640 medium containing 5% FBS (Gibco, USA), 1 mg/mL collagenase I (Biofroxx, Germany), and 0.02 mg/mL DNase I (TIANGEN, China). Tumor tissues were digested for 35 min in cell incubators (Thermo Scientific, USA, 37 °C, 5% CO₂). The digested tissues were ground and filtered through 70-μm cell filters. A Mouse Tumor Infiltrating Lymphocyte Isolation Kit (Solarbio, China) was used to obtain TILs. Cell staining was performed as described above. The gating strategy for the flow cytometric analysis of immune cells in tumor tissue is shown in Additional file 1: Fig. S3.

Zhang Y., Lei Y., Ou Q., Chen M., Tian S., Tang J., Li R., Liang Q., Chen Z, & Wang C. (2024). Listeria-vectored cervical cancer vaccine candidate strains reduce MDSCs via the JAK-STAT signaling pathway. BMC Biology, 22, 88.

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Tumor-Infiltrating Lymphocyte Isolation

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The obtained tumors were enzymatically digested (1.0 mg mL⁻¹ Collagenase IV and 0.2 mg mL⁻¹ DNase I) at 37 °C and homogenized to prepare the corresponding single-cell suspensions. The tumor infiltrated lymphocyte cells were enriched by a Mouse Tumor-infiltrating lymphocyte Isolation Kit (P9000, Solarbio). According to the standard protocols, the single-cell suspensions were stained with the appropriate fluorescent antibodies (Table S1). Finally, the cells were analyzed by flow cytometry.

Chen W., Sheng P., Chen Y., Liang Y., Wu S., Jia L., He X., Zhang C.F., Wang C.Z, & Yuan C.S. (2022). Hypoxia-responsive Immunostimulatory Nanomedicines Synergize with Checkpoint Blockade Immunotherapy for Potentiating Cancer Immunotherapy. Chemical engineering journal (Lausanne, Switzerland : 1996), 451(Pt 2), 138781.

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Isolation of Tumor-Infiltrating Lymphocytes

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Tumors were finely chopped and digested with 1 mg/mL collagenase type V (Sigma) for 1 h at 37°C. After incubation, digested tissues were filtered using a 70 μm cell strainer. The harvested single-cell suspension was incubated with relevant antibody cocktails for flow cytometric assays or utilized to isolate TILs using Mouse Tumor-Infiltrating Lymphocyte Isolation Kit (Solarbio).

Zhang Q., Huang H., Zheng F., Liu H., Qiu F., Chen Y., Liang C.L, & Dai Z. (2020). Resveratrol exerts antitumor effects by downregulating CD8+CD122+ Tregs in murine hepatocellular carcinoma. Oncoimmunology, 9(1), 1829346.

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