Isolation and characterization of tumor-initiating cells from sarcoma patient-derived xenografts

Dan Han, Veronica Rodriguez-Bravo, Sudeh Izadmehr, Josep Domingo-Domenech, Carlos Cordon-Cardo

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The existence and importance of tumor-initiating cells (TICs) have been supported by increasing evidence during the past decade. These TICs have been shown to be responsible for tumor initiation, metastasis, and drug resistance. Therefore, it is important to develop specific TIC-targeting therapy in addition to current chemotherapy strategies, which mostly focus on the bulk of non-TICs. In order to further understand the mechanism behind the malignancy of TICs, we describe a method to isolate and to characterize TICs in human sarcomas. Herein, we show a detailed protocol to generate patient-derived xenografts (PDXs) of human sarcomas and to isolate TICs by fluorescence-activated cell sorting (FACS) using human leukocyte antigen class I (HLA-1) as a negative marker. Also, we describe how to functionally characterize these TICs, including a sphere formation assay and a tumor formation assay, and to induce differentiation along mesenchymal pathways. The isolation and characterization of PDX TICs provide clues for the discovery of potential targeting therapy reagents. Moreover, increasing evidence suggests that this protocol may be further extended to isolate and characterize TICs from other types of human cancers.

Original languageEnglish (US)
Article numbere57011
JournalJournal of Visualized Experiments
Issue number148
StatePublished - Jun 2019


  • Cancer research
  • Human leukocyte antigen-1
  • Human tissue samples
  • Intratumoral heterogeneity
  • Issue 148
  • Patient-derived xenograft
  • Sarcomas
  • Tumor-initiating cells

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology


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