Effective recovery of highly purified CD326+ tumor cells from lavage fluid of patients treated with a novel cell-free and concentrated ascites reinfusion therapy (KM-CART)

Yukino Kimura, Yui Harada, Noriko Yasuda, Takefumi Ishidao, Seiichi Yusa, Keisuke Matsusaki, Yoshikazu Yonemitsu

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Abstract

For the production of tumor-specific vaccines, including dendritic cell (DC) vaccines, the tumor cells themselves are an ideal source. Floating tumor cells in the ascites fluid from patients with malignant ascites are a good candidate source, but it is not easy to obtain pure tumor cells from ascites because of various types of cell contamination as well as protein aggregates. We here report an effective method to recover pure tumor cells from malignant ascites. We used lavage fluid from 13 patients with malignant ascites who were treated with modified cell-free and concentrated ascites reinfusion therapy (KM-CART). Cellular components were separated from the lavage fluid by centrifugation, enzymatic digestion and hemolysis. Tumor cells were purified by depleting CD45+ leukocytes with antibody-conjugated magnetic beads. The tumor cell lysate was extracted by freeze-and-thaw cycles. The mean obtained total cell number was 7.50 × 107 cells (range 4.40 × 106–2.48 × 108 cells). From this fraction, 6.39 × 106 (range 3.23 × 105–2.53 × 107) CD45 cells were collected, and the tumor cell purity was over 80 % defined as CD45CD326+. A sufficient amount of tumor lysate, average  = 2416 μg (range 25–8743 μg), was extracted from CD45CD326+ tumor cells. We here established an effective method to produce highly purified tumor cells from KM-CART lavage fluid. The clinical feasibility of this simple preparation method for generating tumor lysate should be examined in clinical studies of DC vaccines.

Original languageEnglish
Article number780
Pages (from-to)1-6
Number of pages6
JournalSpringerPlus
Volume4
Issue number1
DOIs
Publication statusPublished - Dec 1 2015

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Therapeutic Irrigation
Ascites
Neoplasms
Therapeutics
Cancer Vaccines
Dendritic Cells
Hemolysis
Centrifugation
Digestion
Leukocytes

All Science Journal Classification (ASJC) codes

  • General

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Effective recovery of highly purified CD326+ tumor cells from lavage fluid of patients treated with a novel cell-free and concentrated ascites reinfusion therapy (KM-CART). / Kimura, Yukino; Harada, Yui; Yasuda, Noriko; Ishidao, Takefumi; Yusa, Seiichi; Matsusaki, Keisuke; Yonemitsu, Yoshikazu.

In: SpringerPlus, Vol. 4, No. 1, 780, 01.12.2015, p. 1-6.

Research output: Contribution to journalArticle

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