Isolation of Tumor-Infiltrating Lymphocytes by Ficoll-Paque Density Gradient Centrifugation

  • Yee Sun Tan
  • Yu L. LeiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1960)


With the renewed enthusiasm in immuno-oncology, characterization of the tumor immune microenvironment constitutes an essential and unique aspect to the assessment of therapeutics. The isolation of tumor-infiltrating lymphocytes (TILs) is a desirable approach toward the understanding of antitumor immune response. This chapter provides an effective protocol to mechanically dissociate tumor tissue and generate single-cell suspension from excised tumors. TILs are then isolated by Ficoll-Paque density gradient centrifugation. This protocol is applicable to both human and experimental tumors in immunocompetent murine models.


Tumor-infiltrating lymphocytes Tumor microenvironment Immunotherapy Immune escape Mechanical dissociation 



This work is supported by NIH grants R00 DE024173 (YLL) and R01 DE026728 (YLL), Rogel Cancer Center Fund for Discovery (YLL).


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Periodontics and Oral MedicineUniversity of Michigan School of DentistryAnn ArborUSA
  2. 2.University of Michigan Rogel Cancer CenterAnn ArborUSA
  3. 3.Department of Otolaryngology—Head and Neck SurgeryUniversity of Michigan Health SystemAnn ArborUSA

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