Project: Cell-Diagnostics Sample-Preparation Polymer BioMEMS

BACKGROUND: RARE-CELLS & SILICON BIOSYSTEMS's DEPArray TECHNOLOGY_x000D_Rare cells such as fetal cells in maternal peripheral blood, circulating tumor cells (CTC) or natural therapeutic immune cells have been the subject of increasing research because of their potential in diagnostics, pharmacogenetics and therapeutics. _x000D_Although one can find in scientific literature several demonstrations of feasibility of their retrieval and molecular analysis, tapping their potential in routine clinical practice has been hampered by a lack of reliable, standardized, reproducible and user-friendly methods and tools to handle them._x000D_Silicon Biosystems (SB) has developed a unique proprietary technology (DEPArray), to isolate multiple individual cells in a fairly automated way, which is ideally suited to address this problem._x000D_In the last three years SB has been engaged in the development of protocols for Non-Invasive Prenatal Diagnosis, and more recently expanded its operations to the field of CTC analysis. While the company has demonstrated its potential in this high-added value high-volume markets, it has now understood that one of the critical factors in establishing these protocols is the development of further technologies improving the upstream sample-preparation procedures._x000D_Polymer microfluidic biochips are ideally suited to create low cost disposable which will enable to carry out the required cell preparations. Hence the neeed to work on the concept and, in parallel, on the manufacturing process able to deliver the high volumes and quality required for clinical diagnostics applications._x000D__x000D_PROJECT GOAL: IMPROVE SAMPLE PREPARATION FOR RARE-CELLS ISOLATION WITH DEPArray_x000D_Sample preparation procedures for rare-cell isolation from peripheral-blood samples are currently based on technologies which are unsatisfactory for the goal of implementing into clinical practice Non-Invasive Prenatal Diagnosis based on fetal nucleated Red-Blood-Cells (fetal nRBCs)._x000D_Well established techniques such as centrifugation into density gradients (e.g. Ficoll Hypaque) followed by enrichment of target cells or depletion of unwanted cells with monoclonal antibodies (mAb) coupled to magnetic microbeads typically yield tens to hundred of target cells in a background of few millions of red blood cells (RBC) and few tens of thousands of unwanted nucleated cells._x000D__x000D_Two issues have to be solved to carry out reliably the individual cell selection and sorting with the DEPArray technology:_x000D_1) RBC count is too high, and causes jamming of the DEPArray cages: RBC should be further depleted of at least one order of magnitude._x000D_2) After the enrichment, cell labeling with fluorescent antibodies is needed in order to identify the wanted cells within the DEPArray sorting chamber. Conventional 'tube' protocols -entailing several centrifugation steps- bring about a loss of up to 90% of the cells. Furthermore, cell morphology and fluorescent signal intensity are damaged in delicate cells such as the fetal nRBCs in maternal blood, making it difficult to discriminate them from unwanted cells: a more gentle labeling procedure, decreasing cell loss of at least one order of magnitude is required._x000D__x000D_APPROACH: INNOVATIVE, LOW-COST, POLYMERIC, MICROFLUIDIC BioMEMS_x000D_This project regards innovative microfluidic approaches to solve the aforesaid problems, employing _x000D_*) a common manufacturing technology based on low-cost polymeric devices_x000D_*) a common design approach of fluidic devices based on micromechanical features _x000D__x000D_ANTICIPATED ACHIEVEMENTS:_x000D_We will design, manufacture and validate with cell models and real-world clinical samples innovative microfluidic devices which will enable the introduction of non-invasive prenatal diagnosis into routine clinical use._x000D_We will develop a manufacturing technology based on polymer BioMEMS which significantly advances the state-of-the-art because it combines high-aspect ratio and single-digit micron scale features._x000D_The teechnology will be suitable for the low-cost production of those microfluidic devices and other types of cell-on-chip devices._x000D_

Acronym CellDiaSP (Reference Number: 4809)
Duration 19/08/2009 - 18/08/2013
Project Topic New microfluidic devices for cell sample-preparation are conceived and validated. Unprecedented cell yield and quality enables rare-cell diagnostics. _x000D_ An new single-digit micron-scale, high-aspect ratio manufacturing process for polymer-based BioMEMS is developed to reach a low-cost implementation.
Project Results
(after finalisation)
CO technical achievements are:_x000D_1. The evaluation of the replication of structures with high aspect ratio in polymers_x000D_2. Development of methods for the replication of structures with high aspect ratio of up to 20 by electroplating_x000D_3. Realization of flow cells for the enrichment of cell with higher diameter than erythrocytes_x000D_
Network Eurostars
Call Eurostars Cut-Off 2

Project partner

Number Name Role Country
3 Department of Innovation in Mechanics and Management, University of Padova Partner Italy
3 Mildendo Gesellschaft für mikrofluidische Systeme mbH Partner Germany
3 Silicon Biosystems SpA Coordinator Italy