We performed experiments using PMN-MDSC obtained by two different procedures:
Purification of spleen-derived PMN-MDSC was performed by immunomagnetic positive selection from mice carrying 21-day Lewis lung cell carcinoma tumors with Ly6G microbead kit (Miltenyi RRID: AB_2895065) according to the manufacturer's recommendations.
Differentiated PMN-MDSC obtained from murine bone marrow were generated as previously described (57 (link)). Briefly, mononuclear precursors were obtained from bone marrow of 8- to 12-week-old female C57BL/6J mice (RRID:IMSR_JAX:000664) and cultured in complete medium for 24 hours. Nonadherent cells were recovered 24 hours after extraction and reseeded. These cells were incubated for 7 days with 50% CM obtained from 3-day culture of 8 × 105 genetically modified Lewis lung carcinoma cells and 50% complete medium. Adherent cells were harvested with PBS with 1 mmol/L EDTA and were used for flow cytometry analyses.
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Ruiz-Fernández de Córdoba B., Moreno H., Valencia K., Perurena N., Ruedas P., Walle T., Pezonaga-Torres A., Hinojosa J., Guruceaga E., Pineda-Lucena A., Abengózar-Muela M., Cochonneau D., Zandueta C., Martínez-Canarias S., Teijeira Á., Ajona D., Ortiz-Espinosa S., Morales X., Ortiz de Solórzano C., Santisteban M., Ramos-García L.I., Guembe L., Strnad V., Heymann D., Hervás-Stubbs S., Pío R., Rodríguez-Ruiz M.E., de Andrea C.E., Vicent S., Melero I., Lecanda F, & Martínez-Monge R. (2022). Tumor ENPP1 (CD203a)/Haptoglobin Axis Exploits Myeloid-Derived Suppressor Cells to Promote Post-Radiotherapy Local Recurrence in Breast Cancer. Cancer Discovery, 12(5), 1356-1377.
Corresponding Organization : Clinica Universidad de Navarra
Other organizations :
Unité en Sciences Biologiques et Biotechnologies de Nantes, Nantes Université, Centre National de la Recherche Scientifique, Institut de Cancérologie de l'Ouest, Friedrich-Alexander-Universität Erlangen-Nürnberg, University of Sheffield
Procedure for obtaining PMN-MDSC: 1) Immunomagnetic positive selection from mouse spleens with Ly6G microbead kit, 2) Differentiation from mouse bone marrow precursors with conditioned medium from Lewis lung carcinoma cells
dependent variables
Not explicitly mentioned
control variables
Mouse strain: C57BL/6J
Age of mice: 8- to 12-weeks-old
Sex of mice: Female
controls
Positive control: Not specified
Negative control: Not specified
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