Realtime Monitoring For The Role Of Retinoid On The NB4Cells

Acute promyelocytic leukemia(APL) is a malignant tumor of humanhematopoietic system diseases. It accounts10%-15%for adult acute myeloidleukemia (AML). Because of its rapid onset and progression in the cloned bonemarrow, peripheral blood and other hematopoietic tissue, which can cause death bythe early fatal bleeding and disseminated intravascular coagulation (DIC), the APLcells have attracted much attention the researchers. Thus, the detection of leukemiccells has very important significance in the early diagnosis of leukemia, drugtreatment, post-treatment evaluation. In recent years, the prognosis of APL ischanging, from the worst among AML as it used to be, to currently the best. It hasbeen reported that the application of all-trans-retinoic acid (ATRA) decreases themortality of newly diagnosed patients, which significantly improved the responserate. Therefore, ATRA has been widely accepted and used as a classic treatment forthe APL Diagnosis. Microfluidic chip integrates the General features of biologicaland chemical laboratory into a very small chip by the unique micromachiningtechnique. Due to its size similar to the cell dimension and its controllable inner3Dstructure, the microfluidic chip can make the integrated, systematic cell researchpossible, which enables the cell culture, irritation and tag process on just one chip,and widely applied in the life science field.In this thesis, we fabricated a new microchip for the capture and detection of theNB4cells. We also explored the suitable condition for the cell culture in themicrofluidic chip, and reach the goal of real-time monitoring for the role of retinoidon the NB4cells.1) We first fabricated a new biosensor for the simple and fast Leukemia celldetection. First the gold film was modified with MUA, thus the antibody cancovalently cross-linked on the gold surface. When the leukemia cells flowed throughthe micro-chip, it can be captured by the antibody due to the specificAntibody-Antigen effect. The binding of cell can cause the change of indexrefraction, thus detected by the SPRi technique. And we found there was alogarithmic linear relationship between the signal and cell concentration within acertain range. And the limit of detection (LOD) was61cells per milliliter.2) We also assembled a simple microfluidic chip, to establish a model of cellculture in microchip. The NB4leukemia cells were captured by the antibodies onthe surface of chip, and the cultured in the chip. Then we used retinoic acid tostimulate the leukemia cells, which can cause the leukemia cell change. Thus we can reach the goal of real-time monitoring of drug effect by SPRi, and explore themechanism on cells.