Microfluidic Toolbox for Personalized Medicine – from Ex-vivo Drug Testing of Solid Tumor to Detection and Retrieval of Circulating Tumor Cell
Date Issued
2015
Date
2015
Author(s)
Ma, Wei-Yuan
Abstract
Circulating tumor cell (CTC) now is considered as promising biomarker for cancer diagnosis and prognosis. Many studies had reported the overall survival of patients was related to the number of CTCs detected in whole blood. However, patient-derived CTC count may not enough for doctors to make an actionable decision since CTC possess high heterogeneity, yet the sensitivity of current technologies for CTC enrichment remains to be validated. To further investigate the role of CTC in metastasis process, there has been great interest in subtyping, single cell profiling and molecular analysis of CTC recently. This thesis presents a microfluidic-based, automatic, seamless system enabling single CTC isolation & retrieval from whole blood sample. Cell line results of isolation system showed the ability to recovery CTCs with 89.7 ± 4.7% efficiency from 2ml of human whole blood regardless of their EpCAM expression level. Sensitivity is achieved at low concentrations and recovery of cells through the range of 10 to 300 cells. Results of single cell retrieval system showed the successful retrieval rate reached nearly 100% of target cells and the average retrieval purity was over 90%. On the other hand, for downstream cellular assay on rare cell or limited cell-supplied sample, this thesis presents a microfluidic-based, 96well-formatted plate with built-in micro gap to preserve these precious cells up to 99.2% during multiple assay/wash procedures. Comparing to traditional plate much less cell was needed when using this plate. Drug response profiling on limit cell-supplied tumor sample was performed as one application of this plate. Results on cell line (MCF7 and MDA-MB-231) and patient tumor sample showed the feasibility of cellular analysis in rare cell scenario using this plate. In this thesis, an automatic CTC isolation & retrieval system and a micro-gap plate for cellular analysis of rare cell were presented. Solid evidence of feasibility, stability, and robustness of this whole system were also provided. Conceivably, this microfluidic-based system is applicable to personalized medicine, providing patient-derived information for doctors to tailor therapeutic interventions for patients.
Subjects
Microfluidics
Circulating tumor cells
Single cell retrieving
Chemosensitivity profiling
Persoalized medicine
SDGs
Type
thesis