Mouse nk cell isolation kit

Manufactured by STEMCELL

Sourced in United Kingdom

The Mouse NK Cell Isolation Kit is a laboratory product designed to isolate natural killer (NK) cells from mouse splenocytes. It utilizes a magnetic-based separation technique to enrich for the NK cell population.

Automatically generated - may contain errors

Lab products found in correlation

T25 flask, by Thermo Fisher Scientific (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Hil 15, by Miltenyi Biotec (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Facsaria 3, by BD (1 mentions) Ril 2, by Roche (1 mentions) Human nk cell isolation kit, by STEMCELL (1 mentions) Lymphocyte separation medium, by MP Biomedicals (1 mentions) Cellevent caspase 3 7 green detection reagent, by Thermo Fisher Scientific (1 mentions) Facsaria 2, by BD (1 mentions) Mouse cd8 t cell isolation kit, by STEMCELL (1 mentions) Clone htk888, by BioLegend (1 mentions) Clone 145 2c11, by BioLegend (1 mentions)

6 protocols using mouse nk cell isolation kit

Xenoplantation of HS-MM cells and NK cell therapy

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Xenoplantation of HS-MM cells was performed as previously described (9 (link),10 (link)). Briefly, SCID-Beige mice were injected with 2.5x10⁷ HS-MM cells into the aponeuroses of the thighs, subsequently 9 mice were randomly divided to three groups, of three mice. A total of 0.3 ml saline, with or without 1x10⁴ murine or 1.5x10⁶ human NK cells was intravenously injected to SCID-Beige mice two weeks following HS-MM xenoplantation. A total of 8 weeks following xenoplantation, the mice were sacrificed, and the tumor size was measured using a caliper and the following formula: Volume=(width² x length)/2.
NK cells were harvested from splenic cells of BALB/c mice using a mouse NK cell isolation kit (Stemcell Technologies, Inc.), or collected from peripheral blood samples from a healthy volunteer using negative selection and a NK Cell Isolation kit, MACS system (Miltenyi Biotec, Inc.) according to the manufacturer's protocol. Written informed consent was provided from the healthy volunteer prior to the start of the study. The study was approved by the Committee of Gifu Graduate School of Gifu, Japan (approval no. 27-80, 2020-066).
The isolated human NK cells were cultured in T25 flasks (Thermo Fisher Scientific, Inc.) with 10 ml KBM502 medium (Kohjin-bio, Co., Ltd.) at 37˚C in a humidified incubator with 5% CO₂ for seven days.

Hanamatsu Y., Saigo C., Kito Y, & Takeuchi T. (2020). An obstructive role of NK cells on metastatic growth of clear-cell sarcoma cells in a xenoplant murine model. Molecular and Clinical Oncology, 14(1), 9.

+ Open protocol

+ Expand

Expansion and Characterization of Mouse NK Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

NK cells were purified from mouse spleens using Mouse NK cell isolation Kit according to manufacturer’s specifications (STEMCELL #19855). NK cells were then expanded for 6 days with hIL-15 50 ng/mL (Miltenyi #130-095-765) in RPMI 20% FCS supplemented with L-glutamine (2 mM; Gibco), penicillin/streptomycin (100 µg/mL; Gibco), Sodium Pyruvate (1 mM; Gibco) and finally FACS sorted (NK1.1⁺, NKp46⁺, CD3^-, CD19^-) (BD Biosciences FACSAria III). After overnight starvation in the absence of IL-15, Cish^+/+Ncr1 iCre or Cish^fl/flNcr1^iCre NK cells were stimulated or not during 4 hours with IL-2 (Proleukin, Novartis pharma SAS) or IL-15.

Bernard P.L., Delconte R., Pastor S., Laletin V., Costa Da Silva C., Goubard A., Josselin E., Castellano R., Krug A., Vernerey J., Devillier R., Olive D., Verhoeyen E., Vivier E., Huntington N.D., Nunes J, & Guittard G. (2022). Targeting CISH enhances natural cytotoxicity receptor signaling and reduces NK cell exhaustion to improve solid tumor immunity. Journal for Immunotherapy of Cancer, 10(5), e004244.

+ Open protocol

+ Expand

Isolation and Culture of Human and Murine NK Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human blood samples from normal healthy donors were drawn for research purposes under a protocol approved by the institutional review board with informed consent. PBMCs were isolated by density gradient centrifugation using lymphocyte separation medium (MP Biomedicals, Solon, OH). Human NK cells were purified from PBMCs by negative selection using a human NK cell isolation kit (StemCell Technologies) as described (33 (link)). These cells were 97–99% CD3-CD56+ as assessed by flow cytometry. The human NK cell line NKL was cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS), 1 mM sodium pyruvate, and 200 U/ml recombinant IL-2 (rIL-2) (Roche). K562, 721.221, and KCL-22 cells were cultured in Iscove's modified Dulbecco's medium (IMDM) supplemented with 10% FBS and 2 mM L-glutamine. YAC-1, RMA, and RMA-s cells were cultured in RPMI1640 supplemented with 5% FBS and 2 mM L-glutamine. B16F10 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% FBS and 2 mM L-glutamine. In some experiments, PBMCs were activated with 200 U/ml rIL-2 for 24 h. Male C57BL/6 mice were used at 6–7 weeks old. Murine NK cells were purified from splenocytes by negative selection using a mouse NK cell isolation kit (StemCell Technologies) according to the manufacturer's instructions.

Kwon H.J., Lee H., Choi G.E., Kwon S.J., Song A.Y., Kim S.J., Choi W.S., Hwang S.H., Kim S.C, & Kim H.S. (2018). Ginsenoside F1 Promotes Cytotoxic Activity of NK Cells via Insulin-Like Growth Factor-1-Dependent Mechanism. Frontiers in Immunology, 9, 2785.

+ Open protocol

+ Expand

Isolation of Mouse Natural Killer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Mouse NK Cell Isolation kit was used according to the manufacturer's instructions (19855; Stemcell Technologies UK Ltd.) using Falcon 5-ml polystyrene round-bottom tubes on the EasySep magnet. Splenocyte suspensions at a density of 1×10⁸ cells/ml were centrifuged at 300 × g for 10 min at 4°C and resuspended in 1 ml of recommended medium in a 5-ml polystyrene tube. A total of 50 µl EasySep mouse NK cell isolation cocktail was added, and cells were incubated at room temperature (15–25°C) for 10 min. Subsequently, 100 µl EasySep Streptavidin RapidSpheres 50002 was added, and cells were incubated at room temperature (15–25°C) for 5 min. Cell suspensions were brought up to a total volume of 2.5 ml by adding the recommended medium, and were placed into the magnet for 5 min at room temperature (15–25°C). The EasySep magnet was removed and the desired fraction was poured off into a 15-ml tube three times following 5 min separations in the magnet. The isolated cells in the 15-ml tube were naïve NK cells.

Wang G., Yu G., Wang D., Guo S, & Shan F. (2017). Comparison of the purity and vitality of natural killer cells with different isolation kits. Experimental and Therapeutic Medicine, 13(5), 1875-1883.

+ Open protocol

+ Expand

NK Cell-Mediated Cytotoxicity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

NK cells were sorted from the spleen of leukemic or control mice using mouse NK cell isolation kit (StemCell, 19855) and FACS Aria II (BD Biosciences). Target YAC-1 cells or FLB1 cells were stained with 4µM of Cell Proliferation Dye eFluor™ 670 (Life Technologies) according to manufacturer’s instructions. Sorted NK cells were then incubated with target cells for four hours at 37°C at different effector to target (E:T) ratios. Target cell killing was measured using CellEvent™ Caspase-3/7 Green Detection Reagent (Life technologies) and analyzed by flow cytometry.

Bou-Tayeh B., Laletin V., Salem N., Just-Landi S., Fares J., Leblanc R., Balzano M., Kerdiles Y.M., Bidaut G., Hérault O., Olive D., Aurrand-Lions M., Walzer T., Nunès J.A, & Fauriat C. (2021). Chronic IL-15 Stimulation and Impaired mTOR Signaling and Metabolism in Natural Killer Cells During Acute Myeloid Leukemia. Frontiers in Immunology, 12, 730970.

+ Open protocol

+ Expand

Cytotoxic Degranulation of NK and CD8+ T Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cytotoxic degranulation was assessed by measuring cell surface expression of CD107a as previously described (47 (link)). Resting or IL-2–activated NK cells isolated from splenocytes by negative selection using a mouse NK cell isolation kit (STEMCELL Technologies) were stimulated with YAC-1 or B16F10 cells for 4 hours. Lymphocytes were gated on forward scatter/side scatter, and the CD107a expression of CD3ε⁻NKp46⁺ NK cells was analyzed by flow cytometry. NK cell degranulation was determined by the percent increase of CD107a⁺ NK cells after stimulation with YAC-1 or B16F10 cells relative to CD107a⁺ NK cells without stimulation (ΔCD107a⁺ cells). For assessing CD8⁺ T cell-cytotoxic activity, IL-2–activated CD8⁺ T cells were isolated from splenocytes by negative selection using a mouse CD8⁺ T cell isolation kit (STEMCELL Technologies) and were stimulated with P815 cells preincubated with anti-CD3ε antibody (10 μg/ml, clone 145-2C11; BioLegend) or isotype control antibody (10 μg/ml, clone HTK888; BioLegend) for 4 hours. P815 cells are FcR⁺ and serve as a classical target for T cell cytotoxicity via their ability to bind antibodies to specific activating receptors (for example, CD3ε). In doing so, they can stimulate T cells through CD3ε. Cytotoxic degranulation was measured by surface expression of CD107a on CD3ε⁺CD8a⁺ T cells after lymphocyte gate.

Hyun Y.M., Seo S.U., Choi W.S., Kwon H.J., Kim D.Y., Jeong S., Kang G.Y., Yi E., Kim M., Ryu H.J., Looney M.R., Choi E.Y, & Kim H.S. (2020). Endogenous DEL-1 restrains melanoma lung metastasis by limiting myeloid cell–associated lung inflammation. Science Advances, 6(45), eabc4882.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!