| Background:Hepatocellular carcinoma(HCC)is a common malignant tumor.The average annual incidence of HCC in China was 423,000 with a disappointed 5-year survival rate of 14.1%.Surgery is the most important treatment for HCC,but more than 70%of patients have missed the opportunity for surgery.Therefore,it is of great significance for prolonging the survival time of HCC that exploring new screening strategies for early diagnosis.To date,alpha-fetoprotein(AFP)is still the most commonly used biomarker for HCC screening.Unfortunately,it presents limited sensitivity and specificity.Contrast-enhanced ultrasound,enhanced CT,needle biopsy,etc.are useful for the diagnosis.However,none of these can be widely used as a routine screening method for HCC due to the harmful drugs,X-ray and invasive puncture.CTCs have high sensitivity for the early diagnosis of HCC,and only need to be detected from peripheral blood of patients with the advantage of non-invasiveness,repeated sampling,and the high auxiliary diagnostic value.However,the existing commercial CTCs detection technologies are too expensive and delayed to be applied for HCC screening.Therefore,there is an urgent need to develop a new CTCs detection platform with low cost,simple operation and timely reporting.Objectives:To develop an HCC CTCs detection platform that can be widely used in primary medical units for HCC screening with low cost,simple operation,and timely reporting,so as to improve the early diagnosis rate of HCC.Methods:First,Autodesk Fusion 360 and Comsol were used to simulate pipeline and fluid distribution,and modules for reagent collection and pressure introduction were 3D printed followed the Autodesk Fusion360 designs.Secondly,the chip design was optimized in order to meet the need of CTCs isolation,and accordingly,chip mold was fabricated by deep silicon etching.Then,the performance of the above modules in cell isolation and immune identification was tested by human tumor cell lines to simulate peripheral blood CTCs.Further,a new platform for CTCs detection was established by means of solenoid valve regulation and module assembly.Finally,the platform was used to detect CTCs from HCC patients to analyze the clinical indication of CTCs.Results:(1)Pressure-driven microfluidic introduction was applied to CTCs detection,which gave full play to its advantages of sensitive control,low crosscontamination,and light cell damage.The proposed platform improved the efficiency of reagent exchanging and showed great potential for the analysis of low-abundance cells such as CTCs.(2)Solenoid valves were used for fluid control which reduced equipment hardware costs and maintenance difficulties.Therefore,the platform was suitable for promotion in primary medical institutions with poor technical conditions.(3)Blood cell mass as background was improved in the optimized microfluidic chip.In addition,the immune efficiency for CTCs identification was significantly improved by oscillatory mixing.Compared with static incubation,both antibody and time consumption were saved by 90%.(4)By integrating the above technology platforms,a new platform,Cellcaptor,for automated HCC CTCs detection with independent intellectual property rights has been established,which can complete the CTCs detection process within 90 minutes.(5)The Cellcaptor system was used to detect CTCs from clinically diagnosed HCC patients and healthy donors.The results showed that CTCs could reflect the biological characteristics of HCC.And the detection performance of Cellcaptor was similar to that of other mainstream CTCs detection instruments but with low cost and simple operation.Conclusions:We developed a new CTCs detection platform with low cost,simple operation,timely reporting,and easy maintenance.With the guiding value for HCC diagnosis,prognosis and treatment decision,the platform is expected to be popularized and applied in primary medical units,which will help to improve the early diagnosis rate of HCC,thereby improving the 5-year survival rate. |
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