CIRCULATING TUMOR CELLS, THE SIZE-BASED VIABLE ENRICHMENT AND THE EX VIVO EXPANSION

open_access
Open Access
Author:
Hao, Sijie
Graduate Program:
Bioengineering
Degree:
Doctor of Philosophy
Document Type:
Dissertation
Date of Defense:
September 12, 2017
Committee Members:
Siyang Zheng, Dissertation Advisor/Co-Advisor
William O Hancock, Committee Chair/Co-Chair
Jian Yang, Committee Member
Xiaojun Lian, Committee Member
ANDREA MARIE MASTRO, Outside Member
Keywords:
Cancer metastasis
Bone-on-a-chip
Microfluidics
Circulating tumor cells
CTC Ex Vivo culture
CTC enrichment
Abstract:
Cancer metastases account for 90% of cancer deaths. Circulating tumor cells or CTCs spread from the primary tumor sites to the secondary sites via the circulatory systems and thus promote metastasis. The molecular aberration of CTCs was shown to correlate with the patients’ disease spread and the efficacy of the treatment. In the peripheral blood, however, only a few CTCs are transiently present among millions of nucleated hematopoietic cells. This extreme rarity presents a huge challenge to the downstream characterization of CTCs. Enrichment of viable CTCs could permit functional analyses of CTCs to broaden understanding of metastatic disease. This work first presents a novel Separable Bilayer Microfiltration device for viable, label-free enrichment of circulating tumor cells, followed by viability test of FMSA-enriched CTCs by using a replication-competent recombinant adenovirus driven by a human telomerase gene (hTERT) promoter. We describe the optimization of FMSA 2D layout for semi-automated microscopic scanning of whole-FMSA for CTC detection and morphology documentation. We continue to demonstrate the successful expansion of mouse CTCs via a novel in-situ culture strategy. Moreover, our effort in ex vivo culture of patient CTCs enriched from clinical specimens are also discussed. We report a bone-on-a-chip (BC) microfluidic device for spontaneous growth of a 3D, mineralized, collagenous bone tissue. This device facilitates the easy, frequent observations of cancer cell/bone matrix interaction for in vitro study of cancer bone metastasis.