Abstract
Recently, single cell cloning techniques have been gradually developed benefited from their important roles in monoclonal antibody screening, tumor heterogeneity research fields, etc. In this study, we developed a high throughput device containing 1400 lateral chambers to efficiently isolate single cells and carry out long-term single cell clonal cultivation as well as tumor heterogeneity studies. Most of the isolated single cells could proliferate normally nearly as long as three weeks and hundreds of clones could be formed once with one device, which made it possible to study tumor heterogeneity at single cell level. The device was further used to examine tumor heterogeneity such as morphology, growth rate, anti-cancer drug tolerance as well as adenosine triphosphate-binding cassette (ABC) transporter ABCG2 protein expression level. Except for the single cell isolation and tumor heterogeneity studies, the device is expected to be used as an excellent platform for drug screening, tumor biomarker discovering and tumor metastasis assay.
Highlights
Tumor cell heterogeneity on cell morphology, growth rate, drug sensitivity and ABCG2 protein expression difference have been conducted at single cell clone level on the device
COMSOL Multiphysics 4.3 simulation result showed that the surface velocity within the culture chambers was extremely low, which indicated that the impact of hydrodynamic shear stress was negligible, due to a positive correlation between hydrodynamic shear stress and flow rate[32]
Dynamic perfusion system is essential for fresh medium supplement and poisonous metabolites discharging based on the effective diffusion, which is important for long-term clonal cultivation
Summary
Aside from the physical structures mentioned above, several other similar models have emerged[20,21] Some of these models provided novel approaches for single cell isolation, some of the designs based on physical constraints inevitably exposed the isolated single cells under hydrodynamic shear stress which had detrimental impacts on isolated cells and made the relevant device not apt for long-term clonal cultivation. Based on the recent researches as well as our studies, there are three key points essential for long-term single cell clonal cultivation in microfluidic devices: 1) adequate space for single cell long-term clonal proliferation; 2) dynamic perfusion system for fresh nutrient supplement and poisonous metabolite excretion; 3) minimum negative impacts of hydrodynamic shear stress on isolated cells. Tumor cell heterogeneity on cell morphology, growth rate, drug sensitivity and ABCG2 protein expression difference have been conducted at single cell clone level on the device
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call