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仅通过血液过滤分离循环肿瘤细胞的新型滤器的验证。

Verification of A New Filter for Isolation of Circulating Tumor Cells by Only Blood Filtration.

机构信息

Department of Diagnostic Pathology, Nara Medical University School of Medicine, Kashihara, Japan.

Department of Diagnostic Pathology, Nara Prefecture General Medical Center, Nara, Japan.

出版信息

Anticancer Res. 2022 Sep;42(9):4305-4310. doi: 10.21873/anticanres.15930.

Abstract

BACKGROUND/AIM: Since circulating tumor cells (CTCs) are precursors of metastatic lesions, extracting CTCs from whole blood is useful in obtaining information for cancer treatment. One of the CTC isolation methods is the size selection method; however, since the conventional methods are expensive and cumbersome, we developed an affordable and simple filter, whose usefulness is verified in this study.

MATERIALS AND METHODS

The new filter [hereafter, soft micropore filter (S-MPF)] is made up of a polyethylene film with a thickness of 15 μm and conical pores having a diameter of 8-10 μm, which are opened uniformly (opening rate, 20%). This filter can filter whole blood by free-falling under gravity. The possibilities of the filter's usage for model CTC isolation, immunostaining, short-term cell culture, and gene mutation detection in extracted model CTCs were verified.

RESULTS

S-MPF was able to extract model CTCs with an isolation rate of up to 15%. These model CTCs were detected by cytology, immunostaining, and culture by short-term incubation of filtered cells. Furthermore, genetic mutations were identified in the cultured cells. In addition, CTC isolation from the peripheral blood of patients with lung cancer was demonstrated by setting the volume of collected blood to 15 ml to prevent a low recovery rate.

CONCLUSION

The S-MPF can be used to extract model CTCs quickly and easily. Moreover, cytological diagnosis, immunostaining, short-term culture, and gene mutation search are possible with this filter. Given its proven applicability in clinical samples, this filter can be used in clinical settings.

摘要

背景/目的:循环肿瘤细胞(CTC)是转移灶的前体,因此从全血中提取 CTC 有助于获取癌症治疗信息。CTC 分离方法之一是大小选择法;然而,由于传统方法昂贵且繁琐,我们开发了一种经济实惠且简单的过滤器,在本研究中验证了其有用性。

材料和方法

新型过滤器[以下简称软微孔过滤器(S-MPF)]由厚度为 15μm 的聚乙烯膜和直径为 8-10μm 的锥形孔组成,这些孔均匀地打开(开口率为 20%)。该过滤器可以通过重力自由下落过滤全血。验证了该过滤器用于模型 CTC 分离、免疫染色、提取模型 CTC 中的短期细胞培养和基因突变检测的可能性。

结果

S-MPF 能够提取高达 15%的模型 CTC。通过过滤细胞的短期孵育进行细胞学、免疫染色和培养检测到这些模型 CTC。此外,还鉴定了培养细胞中的基因突变。此外,通过将采集的血液量设置为 15ml 来防止回收率低,从肺癌患者的外周血中分离出 CTC 得到了证实。

结论

S-MPF 可用于快速轻松地提取模型 CTC。此外,该过滤器还可以进行细胞学诊断、免疫染色、短期培养和基因突变搜索。鉴于其在临床样本中的适用性已得到证实,该过滤器可在临床环境中使用。

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