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High depletion of breast cancer cells from the peripheral blood with the method of non-specific separation

机译:非特异性分离法从外周血中大量清除乳腺癌细胞

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摘要

Circulating epithelial tumour cells (CETCs) play an important role in the formation of metastases in breast cancer patients. The depletion of such CETCs from peripheral blood of breast cancer patients using non-specific separation (without antibodies) of tumour cells from normal blood leucocytes might contribute to reduce the load of the patient’s blood with tumour cells and subsequently reduce the probability of metastasis formation. This method is based on cell type-specific interaction of living cells with Carboxymethyl Dextrane (CMD) coated magnetic nanoparticles. We have developed a mild flow separation method using CMD-coated magnetic nanoparticles (core size ca. 25 nm) along with a low-field gradient magnetic separator and an external separation column (blood bag). The ability of tumour cells to preferentially bind such particles and to separate tumour cells from the white blood cells from blood samples of 25 breast cancer patients (fresh and 24-hour stored blood samples) were tested. The circulating tumour cells were quantified before and after separation by maintrac analysis. We achieved a very high depletion rate of tumour cells to < 3% remaining in the investigated 24 hours stored blood samples and ≤14% in all fresh blood samples concurrent with maintaining 56% ± 4% of vital leukocytes in all fresh blood samples.
机译:循环上皮肿瘤细胞(CETC)在乳腺癌患者转移的形成中起重要作用。使用非特异性分离正常血液白细胞的肿瘤细胞(不含抗体)从乳腺癌患者外周血中消耗此类CETC可能有助于减少患者血液中肿瘤细胞的负荷,从而降低转移形成的可能性。此方法基于活细胞与羧甲基右旋糖酐(CMD)包覆的磁性纳米粒子的细胞类型特异性相互作用。我们已经开发了一种温和的流分离方法,该方法使用CMD涂层磁性纳米颗粒(核心尺寸约25 nm),低场梯度磁性分离器和外部分离柱(血袋)进行。测试了肿瘤细胞优先结合这些颗粒并将肿瘤细胞与白细胞分离的能力,该能力来自25名乳腺癌患者的血液样品(新鲜和24小时储存的血液样品)。通过主链分析在分离前后对循环中的肿瘤细胞进行定量。我们实现了非常高的肿瘤细胞耗竭率,在所调查的24小时存储的血液样本中,肿瘤细胞的残存率<3%,在所有新鲜血液样本中≤14%,同时在所有新鲜血液样本中维持56%±4%的重要白细胞。

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