Abstract 418: Ex vivoisolation and culture of circulating tumor cells using microfluidics chip

Abstract

Abstract Background: Circulating tumor cells (CTCs) are the tumor cells shed from primary tumor, which enter the bloodstream and travel to distant anatomic sites to form metastasis. CTCs have been identified as blood-based biomarkers to guide cancer diagnosis and prognosis in many reports. The huge challenge which limits the clinical utility of CTCs, especially in early stage cancer patients, is the rarity. Specifically, there are only 1-10 CTCs in one milliliter of whole blood. Methods: A microfluidic CTC capture device was used for capture and analysis of CTCs from gastric cancer patients. The patients’ samples were referred to the regulation of ethnic commission of Shenzhen People’s Hospital and informed parental consent for the study. CTCs were collected by cells adherent culture using the CCC medium. Immunofluorescence, karyotyping and mude mouse tumorigenesis were used to verify the CTCs. Results: A epithelial phenotype cell cluster which named S-CTC came out in the 24-well plate after 8 days. S-CTC cells can steadily formed clones and showed stem-cell like properties. Positive rate of CD133 were reached to 4.7% which demonstrated by flow cytometry. Immunofluorescence identification of S-CTC were DAPI-positive, Cytokeratin-positive, and CD45-negative. The karyotyping results indicated that the chromosomes were aneuploidthe, the chromosome number of S-CTC varied from 55 to 63. Also S-CTC presented a larger nuclear/cytoplasmic ratio and limitless replicative potential. Furthermore, the S-CTC had huge sphere formation ability, the tumorigenesis of S-CTC cells can be happened by xenografting in immunodeficient mice. Conclusion: A cell S-CTC were successfully isolated and cultured by the technology microfluidic chip. S-CTC was identified using a series of analysis and can be confirmed to be the tumor cell, which would be provide more drug sensitivity information of the patient and open up novel avenues towards the understanding of gastric cancer. Note: This abstract was not presented at the meeting. Citation Format: Huirong Zhang, Pan Zhao, Han Wu, Chang Zou. Ex vivoisolation and culture of circulating tumor cells using microfluidics chip [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 418.