Tumor Biology

, Volume 36, Issue 4, pp 2299–2307 | Cite as

Comparative study of different procedures for the separation of peripheral blood mononuclear cells in cytokine-induced killer cell immunotherapy for hepatocarcinoma

  • Hui Liu
  • Jianyu Li
  • Fengmei Wang
  • Yingtang Gao
  • Ying Luo
  • Peng Wang
  • Chenglong Li
  • Zhengyan Zhu
Research Article


Cytokine-induced killer (CIK) cell immunotherapy exhibits significant advantages in the clinical treatment of tumors. This study was designed to compare the biological characteristics of autologous CIK cells from patients with hepatocarcinoma following different procedures for the separation of peripheral blood mononuclear cells (PBMCs). Forty-four hepatocarcinoma patients were enrolled and distributed into two groups. PBMCs were isolated either using a blood cell separator (apheresis method) or Ficoll lymphocyte separation medium (Ficoll method). The total amount, collection efficacy, and cell status of PBMCs in the two groups were determined. According to the number and status of collected PBMCs, different cultivation procedures were used for their amplification and activation and the proliferation ability, phenotype, and killing activity of CIK cells in the two groups were evaluated. Our results indicated that the number of collected PBMCs in the apheresis group was far more than that in the Ficoll group. However, the isolation rate was lower, and more cellular debris was observed in the apheresis group, which may be the cause of some untoward effects. Following in vitro culture, the enrichment time of CIK cells was longer in the Ficoll group, and the percentages of CD3+CD4+ (Th) and CD4+CD25+ (Treg) cells were higher. In the apheresis group, the percentages of CD3CD56+ (NK) and CD3+CD56+ (NKT) cells were higher, and the CIK cells exhibited a higher cytolytic activity against HepG2 hepatoma cells. In conclusion, different procedures for PBMCs separation can influence the biological activities of CIK cells, and the apheresis method is more effective at enhancing the antitumor efficacy of CIK cells. However, significant attention should be paid to the possibility of adverse reactions in apheresis donors.


Cytokine-induced killer cell Hepatocarcinoma Peripheral blood mononuclear cells Separation modality Antitumor efficacy 



This work was supported by grants from the Tianjin Science Foundation of China (No. 13JCYBJC22500) and the Tianjin Public Health Bureau Science Foundation of China (No. 2010KY03).

Conflicts of interest



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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Hui Liu
    • 1
  • Jianyu Li
    • 2
  • Fengmei Wang
    • 1
  • Yingtang Gao
    • 1
  • Ying Luo
    • 1
  • Peng Wang
    • 1
  • Chenglong Li
    • 1
  • Zhengyan Zhu
    • 1
  1. 1.Key Laboratory of Artificial CellTianjin Third Central HospitalTianjinChina
  2. 2.Logistics College of People’s Armed Police ForcesTianjinChina

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