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Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples

Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small... DE GRUYTER Current Directions in Biomedical Engineering 2020;6(3): 20203035 Stephan Behrens*, Frank Sonntag, Florian Schmieder Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples Abstract: Within the human blood immune cells assume 1 Extraction of immune cells specific functions to modulate and control the human immune response. Consequently, they are used for the diagnosis and treatment of autoimmune and hematologic diseases. Traceless 1.1 Motivation Affinity Cell Selection Technology (Fab-TACS® technology) is a new method for selective isolation of specific immune The human immune system is composed of different cell cells from blood samples. Currently, the technology is used in populations, which develop from a common stem cell by a 60 mL syringe-like cartridge, which is operated by a control cellular differentiation. Within the human blood the fraction of unit (Fabian®). Nevertheless, diagnostic applications require immune cells is called peripheral blood mononuclear cells the reduction of the sample volume to several microliters. (PBMC) including lymphocytes (T cells, B cells, NK cells), Therefore, we developed a cartridge for the purification of monocytes and granulocytes. They all assume specific small blood volumes with the aid of suitable scaling and functions to modulate the human immune response. simulation methods. By implementing a multi domain Consequently, they are used for the diagnosis and treatment of physical systems model of the established cartridge we could autoimmune and hematologic diseases [1]. Nowadays the simulate the separation process. Based on that a microfluidic targeted use of specific immune cells is becoming increasingly cartridge was designed and several prototypes were important for the adoptive transfer of T cells (ACT) as a new manufactured. To operate the microfluidic cartridge a setup field of transfusion medicine. This includes the infusion of and software for fluidic control were created. Afterwards the lymphocytes to mediate anti-tumor, anti-viral or anti- prototype cartridge and the microfluidic control were used to inflammatory effects with the aim of combating diseases such isolate lymphocytes from a buffy coat. By flow cytometric as multiple myeloma [2]. Thus, new technologies are arising analysis we could show that it is possible to isolate to isolate such cell populations from whole blood. Up to now lymphocytes with a purity that is comparable to the established complex multi-stage procedures are required to specifically 60 mL syringe-like cartridge. select these from the cell mixture of the blood. Traceless Affinity Cell Selection Technology (Fab-TACS® technology) Keywords: TACS, microfluidic, in-vitro diagnostics, is a new method for the selective, gentle and highly pure simulation, Cell separation isolation of specific immune cells from whole blood or buffy coat. Currently, the technology is used in a 60 mL syringe-like https://doi.org/10.1515/cdbme-2020-3035 cartridge which is operated by a control unit (Fabian®). This fits well to ACT applications. Nevertheless, for applications in laboratory diagnostics or pediatric oncology the amount of blood is limited to less than 100 µL. Thus TACS® technology has to be transferred to a microfluidic cartridge that ensures purification results similar to the established cartridge. 1.2 TACS technology ______ *Corresponding author: Stephan Behrens: Fraunhofer Institute In contrast to conventional methods, non-magnetic cell for Material and Beam Technology IWS, Dresden, Germany, E-Mail: stephan.behrens@iws.fraunhofer.de affinity chromatography, or TACS® technology (traceless Frank Sonntag, Florian Schmieder: Fraunhofer Institute for affinity cell selection), does not use fluorescent markers or Material and Beam Technology IWS, Dresden, Germany Open Access. © 2020 Stephan Behrens et al., published by De Gruyter. This work is licensed under the Creative Commons Attribution 4.0 License. Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples — 2 magnetic beads, which cause long-term damage to the target released, and they separate from the antigens of the cells and cells [3]. Instead it is based on affinity purification with provide residue-free target cells (F). weakly binding antibodies [4]. The method uses the effect of low but numerous binding cell-specific antibodies, also called avidity effect. These so-called Fab-fragments are those 2 Simulation aided prototyping elements of an antibody that only contain the binding region. They are produced recombinantly with an affinity tag, the Strep-Tag®. The Strep-Tag® is capable of binding to another 2.1 Approach specific protein, Strep-Tactin®. At the beginning of the purification process, the Strep-Tactin® is placed on the To transfer biochemical processes, like the Fab-TACS® agarose matrix (compare figure 1). technology, from a high-volume cartridge into microfluidic applications model based approaches have become indispensable [5]. These include several steps (compare figure 2). The first step is to establish a multi domain physical systems model. The technical processes that are implemented in the model are determined by measuring relevant process parameters. Based on these data the separation process was simulated. Within the next step simulation data were used to specify the design of the novel microfluidic cartridge. Afterwards serval prototypes were manufactured by layer object manufacturing [6] and the performance of the cell selection was characterized. Figure 1: Process flow of the Fab-TACS® technology: 1) Fab- antibody; 2) Strep-Tag®; 3) Strep-Tactin®; 4) Agarose bead; 5) Target cells; 6) Non-target cells; 7) Biotin. A, B - Fab loading; C - Blood loading with connection of target cell with the Fab-antibody; D - Wash step; E - Biotin loading; F - Target cell elution. The illustration is purely functional and do not completely represent the real dimensions. Figure 2: Step by step downscaling process If the Fab-Strep antibodies are now added and bind to the matrix (A, B). Afterwards whole blood is loaded into the 2.2 Modelling and simulation of the matrix. Those cells that have expressed the specific antigen on established cartridge the cell surface are bound within the matrix. The remaining unbound blood cells are rinsed from the column by a washing For the modelling of the established cartridge the following procedure (C, D). Finally, biotin, also known as vitamin H, is systematic methodology has been established for a structured added. It acts as a competitor and releases the binding between approach: Strep-Tag® and Strep-Tactin® and attaches itself irreversibly 1. definition of the subsystems to the matrix (E). Besides its attachment, biotin causes the 2. definition of the interfaces target cells to detach from the column particles. By 3. reduction to simple elements conformational change of the Fab-fragments the binding is 4. compiling key effects 5. model notation Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples — 3 In the first step of modelling a multi domain physical systems cartridge. Thus, the whole setup can be dived into the model of the entire system, including the control unit Fabian® microfluidic application and the control unit that are shown in and the disposable cartridge, was established by using figure 3. For valve switching, the pneumatic control unit (4), SimulationX® (ESI ITI GmbH, Germany). Therefore, logical which can switch between defined overpressure and vacuum, subunits of the system were created and correlated to was first connected to the inputs of the chip (1). The syringe corresponding predefined model equivalents of the software pump (5) was connected to the output of the microfluidic (e.g. syringe like cartridge is modelled as plunger cylinder). cartridge. To control the operation of the pneumatic control Afterwards the interconnection of these system components unit, the syringe pump and the downstream two-way valve (6) was described by specific predefined interfaces (e.g. cartridge a graphical user interface was developed and implemented (7). and control unit are connected by mechanical force). Physical Thus, it was possible to switch between the output storage properties of the model were determined by tribology tanks for the negative fractions (2) and the isolated cells (3). measurements of friction forces and the resistance above the agarose matrix during the separation process in the established cartridge. By simulating the separation process within the physically determined cartridge the system pressure as a major influence for the separation process could be extracted from the model. Subsequently a multi domain physical model of the microfluidic cartridge was generated using a model library for microfluidic applications [7]. 2.3 Design and prototyping of a microfluidic cartridge The basic design properties of the microfluidic layout were extracted from the model of the microfluidic cartridge and its sub models from the microfluidic library. This included the length of the channels as well as the size of the separation camber. Within the downscaling process the size and geometry of the strep activated agarose compartment was the most crucial part as acts as the major binding site for the cells. The volume of the chamber was calculated by an allometric scaling approach [8] to fit the requirements of the affinity binding of cells to the matrix. Nevertheless, the geometric dimensions that fit this volume were determined mostly by microfluidic Figure 3: Schematic illustration of the developed flow control mixing processes. To ensure that sample and buffer media did setup and the implemented setup below: 1) Microfluidic not mix during the separation process, pneumatic control cartridge; 2) Waste tank; 3) Storage of the isolated target cells; 4) pneumatic control unit (PCU); 5) Syring pump; 6) valves were integrated that regulate the opening and closing of Changeover valve; 7) Pump controler. sample and buffer tanks. After the design was finished several prototype cartridges were manufactured using layer object manufacturing technology [6]. 4 Performance test 3 Development of the flow The selectivity of the manufactured prototypes was characterized by isolating lymphocytes from a buffy coat control setup (acquired from DRK Spendeservice, Institute of Transfusion Medicine Dresden). Sixty microliters of the buffy coat were To run the separation process within the microfluidic pipetted into the inlet of three cartridge prototypes. Buffers and cartridges a flow control platform has been developed. It samples were drawn by the syringe pump through the combines a pneumatic control unit (PCU) and a syringe pump cartridges according to the regimen specified by the pneumatic for the regulation of media flow within the microfluidic Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples — 4 control unit. The eluate of the last rinsing step, after the high-volume cartridge. Further developments have to address addition of the elution buffer, was collected separately for each the repeatability and technologies for mass production of the cartridge. These samples and the initial buffy coat were microfluidic cartridge to transfer the technology to point of characterised by flow cytometric analysis to determine the care applications. distribution of PBMCs and erythrocytes. To distinguish between nucleated PBMCs and erythrocytes, cells were Author Statement stained with Höchst 33342 (compare figure 4). The diagrams The authors would like to express great appreciation for the financial support to the German Federal Ministry of Education and Research (BMBF) for the project “SIMPLE-IVD” within the “Innovations for Tomorrow’s Production, Services, and Work” program (FK 02P18C100) implemented by the Project Management Agency Karlsruhe (PTKA). References Figure 4: Scatter plots of single cell signals of Höchst 33342 [1] Kaufmann SHE. Basiswissen Immunologie. Berlin: Springer (H33342) vs. foreward scatter(FSC): A) Buffy coat; A) Purified sample [2] Ali SA, Shi V, Maric I, et al. T cells expressing an anti-B-cell maturation antigen chimeric antigen receptor cause show the gated populations in red, green and violet remissions of multiple myeloma. Blood 2016; 128: 1688– representing the lymphocytes, granulocytes and monocytes. The black gate shows the number of erythrocytes. Compared [3] Grützkau A, Radbruch A. Small but mighty: how the MACS- to the buffy coat the distribution of cell populations after technology based on nanosized superparamagnetic purification showed a significant increase in the proportion of particles has helped to analyze the immune system within lymphocytes in the sample with a mean of 80,1% (compare the last 20 years. Cytometry. Part A : the journal of the table 1) and a maximum value of 93.2 %. Thus, the selectivity International Society for Analytical Cytology 2010; 77: 643– of the microfluidic cartridge is comparable to the established TACS cartridge (selectivity > 90%) [9], which was the initial [4] Weiss R, Gerdes W, Leonhardt F, Berthold R, Sack U, benchmark. Nevertheless, the repeatability of the experiments Grahnert A. A comparative study of two separation methods has to be further improved by geometric changes of the to isolate monocytes. Cytometry. Part A : the journal of the cartridge outlet to minimize erythrocyte accumulation during International Society for Analytical Cytology 2019; 95: 234– the rinsing of the negative fraction. [5] Laurien E, Oertel H. Numerische Strömungsmechanik. 5th Table 1: Mean amount of selected cells from three microfluidic ed. Wiesbaden: Springer Vieweg 2013. cartridge tests: L – lymphocytes; E – erythrocytes; M – monocytes; [6] Sonntag F, Grünzner S, Schmieder F, Busek M, Klotzbach G - granulocytes U, Franke V. Multilayer based lab-on-a-chip-systems for L [%] E [%] M [%] G [%] substance testing. In: Klotzbach U, Washio K, Arnold CB, Buffy coat 38,7 55,4 2,5 3,6 editors. Laser-based Micro- and Nanoprocessing IX: SPIE; Microfluidic 2015. 93510C (SPIE Proceedings). 80,1 ± 10,01 19,7 ± 10,0 0,0 ± 0,01 0,1 ± 0,1 cartridge [7] Busek M. Modellierung und Regelung der Nährstoffversorgung in mikrofluidischen Zellkultursystemen. Dresden: TUDpress 2018. 5 Conclusion [8] Sung JH, Wang Y, Shuler ML. Strategies for using mathematical modeling approaches to design and interpret This work shows that simulation based down scaling is a multi-organ microphysiological systems (MPS). APL suitable method to transfer a medium volume cartridge for bioengineering 2019; 3: 21501. blood cell separation from millilitre to microliter scale. The [9] IBA GmbH. Isolation Kit forCD3+ cellsfrom Whole Blood prototype of the microfluidic cartridge yields to similar purity (human)or Buffy Coat (human) with FABian®. in the isolation of lymphocytes compared to the established http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Current Directions in Biomedical Engineering de Gruyter

Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples

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Publisher
de Gruyter
Copyright
© 2020 by Walter de Gruyter Berlin/Boston
eISSN
2364-5504
DOI
10.1515/cdbme-2020-3035
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Abstract

DE GRUYTER Current Directions in Biomedical Engineering 2020;6(3): 20203035 Stephan Behrens*, Frank Sonntag, Florian Schmieder Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples Abstract: Within the human blood immune cells assume 1 Extraction of immune cells specific functions to modulate and control the human immune response. Consequently, they are used for the diagnosis and treatment of autoimmune and hematologic diseases. Traceless 1.1 Motivation Affinity Cell Selection Technology (Fab-TACS® technology) is a new method for selective isolation of specific immune The human immune system is composed of different cell cells from blood samples. Currently, the technology is used in populations, which develop from a common stem cell by a 60 mL syringe-like cartridge, which is operated by a control cellular differentiation. Within the human blood the fraction of unit (Fabian®). Nevertheless, diagnostic applications require immune cells is called peripheral blood mononuclear cells the reduction of the sample volume to several microliters. (PBMC) including lymphocytes (T cells, B cells, NK cells), Therefore, we developed a cartridge for the purification of monocytes and granulocytes. They all assume specific small blood volumes with the aid of suitable scaling and functions to modulate the human immune response. simulation methods. By implementing a multi domain Consequently, they are used for the diagnosis and treatment of physical systems model of the established cartridge we could autoimmune and hematologic diseases [1]. Nowadays the simulate the separation process. Based on that a microfluidic targeted use of specific immune cells is becoming increasingly cartridge was designed and several prototypes were important for the adoptive transfer of T cells (ACT) as a new manufactured. To operate the microfluidic cartridge a setup field of transfusion medicine. This includes the infusion of and software for fluidic control were created. Afterwards the lymphocytes to mediate anti-tumor, anti-viral or anti- prototype cartridge and the microfluidic control were used to inflammatory effects with the aim of combating diseases such isolate lymphocytes from a buffy coat. By flow cytometric as multiple myeloma [2]. Thus, new technologies are arising analysis we could show that it is possible to isolate to isolate such cell populations from whole blood. Up to now lymphocytes with a purity that is comparable to the established complex multi-stage procedures are required to specifically 60 mL syringe-like cartridge. select these from the cell mixture of the blood. Traceless Affinity Cell Selection Technology (Fab-TACS® technology) Keywords: TACS, microfluidic, in-vitro diagnostics, is a new method for the selective, gentle and highly pure simulation, Cell separation isolation of specific immune cells from whole blood or buffy coat. Currently, the technology is used in a 60 mL syringe-like https://doi.org/10.1515/cdbme-2020-3035 cartridge which is operated by a control unit (Fabian®). This fits well to ACT applications. Nevertheless, for applications in laboratory diagnostics or pediatric oncology the amount of blood is limited to less than 100 µL. Thus TACS® technology has to be transferred to a microfluidic cartridge that ensures purification results similar to the established cartridge. 1.2 TACS technology ______ *Corresponding author: Stephan Behrens: Fraunhofer Institute In contrast to conventional methods, non-magnetic cell for Material and Beam Technology IWS, Dresden, Germany, E-Mail: stephan.behrens@iws.fraunhofer.de affinity chromatography, or TACS® technology (traceless Frank Sonntag, Florian Schmieder: Fraunhofer Institute for affinity cell selection), does not use fluorescent markers or Material and Beam Technology IWS, Dresden, Germany Open Access. © 2020 Stephan Behrens et al., published by De Gruyter. This work is licensed under the Creative Commons Attribution 4.0 License. Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples — 2 magnetic beads, which cause long-term damage to the target released, and they separate from the antigens of the cells and cells [3]. Instead it is based on affinity purification with provide residue-free target cells (F). weakly binding antibodies [4]. The method uses the effect of low but numerous binding cell-specific antibodies, also called avidity effect. These so-called Fab-fragments are those 2 Simulation aided prototyping elements of an antibody that only contain the binding region. They are produced recombinantly with an affinity tag, the Strep-Tag®. The Strep-Tag® is capable of binding to another 2.1 Approach specific protein, Strep-Tactin®. At the beginning of the purification process, the Strep-Tactin® is placed on the To transfer biochemical processes, like the Fab-TACS® agarose matrix (compare figure 1). technology, from a high-volume cartridge into microfluidic applications model based approaches have become indispensable [5]. These include several steps (compare figure 2). The first step is to establish a multi domain physical systems model. The technical processes that are implemented in the model are determined by measuring relevant process parameters. Based on these data the separation process was simulated. Within the next step simulation data were used to specify the design of the novel microfluidic cartridge. Afterwards serval prototypes were manufactured by layer object manufacturing [6] and the performance of the cell selection was characterized. Figure 1: Process flow of the Fab-TACS® technology: 1) Fab- antibody; 2) Strep-Tag®; 3) Strep-Tactin®; 4) Agarose bead; 5) Target cells; 6) Non-target cells; 7) Biotin. A, B - Fab loading; C - Blood loading with connection of target cell with the Fab-antibody; D - Wash step; E - Biotin loading; F - Target cell elution. The illustration is purely functional and do not completely represent the real dimensions. Figure 2: Step by step downscaling process If the Fab-Strep antibodies are now added and bind to the matrix (A, B). Afterwards whole blood is loaded into the 2.2 Modelling and simulation of the matrix. Those cells that have expressed the specific antigen on established cartridge the cell surface are bound within the matrix. The remaining unbound blood cells are rinsed from the column by a washing For the modelling of the established cartridge the following procedure (C, D). Finally, biotin, also known as vitamin H, is systematic methodology has been established for a structured added. It acts as a competitor and releases the binding between approach: Strep-Tag® and Strep-Tactin® and attaches itself irreversibly 1. definition of the subsystems to the matrix (E). Besides its attachment, biotin causes the 2. definition of the interfaces target cells to detach from the column particles. By 3. reduction to simple elements conformational change of the Fab-fragments the binding is 4. compiling key effects 5. model notation Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples — 3 In the first step of modelling a multi domain physical systems cartridge. Thus, the whole setup can be dived into the model of the entire system, including the control unit Fabian® microfluidic application and the control unit that are shown in and the disposable cartridge, was established by using figure 3. For valve switching, the pneumatic control unit (4), SimulationX® (ESI ITI GmbH, Germany). Therefore, logical which can switch between defined overpressure and vacuum, subunits of the system were created and correlated to was first connected to the inputs of the chip (1). The syringe corresponding predefined model equivalents of the software pump (5) was connected to the output of the microfluidic (e.g. syringe like cartridge is modelled as plunger cylinder). cartridge. To control the operation of the pneumatic control Afterwards the interconnection of these system components unit, the syringe pump and the downstream two-way valve (6) was described by specific predefined interfaces (e.g. cartridge a graphical user interface was developed and implemented (7). and control unit are connected by mechanical force). Physical Thus, it was possible to switch between the output storage properties of the model were determined by tribology tanks for the negative fractions (2) and the isolated cells (3). measurements of friction forces and the resistance above the agarose matrix during the separation process in the established cartridge. By simulating the separation process within the physically determined cartridge the system pressure as a major influence for the separation process could be extracted from the model. Subsequently a multi domain physical model of the microfluidic cartridge was generated using a model library for microfluidic applications [7]. 2.3 Design and prototyping of a microfluidic cartridge The basic design properties of the microfluidic layout were extracted from the model of the microfluidic cartridge and its sub models from the microfluidic library. This included the length of the channels as well as the size of the separation camber. Within the downscaling process the size and geometry of the strep activated agarose compartment was the most crucial part as acts as the major binding site for the cells. The volume of the chamber was calculated by an allometric scaling approach [8] to fit the requirements of the affinity binding of cells to the matrix. Nevertheless, the geometric dimensions that fit this volume were determined mostly by microfluidic Figure 3: Schematic illustration of the developed flow control mixing processes. To ensure that sample and buffer media did setup and the implemented setup below: 1) Microfluidic not mix during the separation process, pneumatic control cartridge; 2) Waste tank; 3) Storage of the isolated target cells; 4) pneumatic control unit (PCU); 5) Syring pump; 6) valves were integrated that regulate the opening and closing of Changeover valve; 7) Pump controler. sample and buffer tanks. After the design was finished several prototype cartridges were manufactured using layer object manufacturing technology [6]. 4 Performance test 3 Development of the flow The selectivity of the manufactured prototypes was characterized by isolating lymphocytes from a buffy coat control setup (acquired from DRK Spendeservice, Institute of Transfusion Medicine Dresden). Sixty microliters of the buffy coat were To run the separation process within the microfluidic pipetted into the inlet of three cartridge prototypes. Buffers and cartridges a flow control platform has been developed. It samples were drawn by the syringe pump through the combines a pneumatic control unit (PCU) and a syringe pump cartridges according to the regimen specified by the pneumatic for the regulation of media flow within the microfluidic Modelling and prototyping of a microfluidic cartridge for extracting immune cells from small volume blood samples — 4 control unit. The eluate of the last rinsing step, after the high-volume cartridge. Further developments have to address addition of the elution buffer, was collected separately for each the repeatability and technologies for mass production of the cartridge. These samples and the initial buffy coat were microfluidic cartridge to transfer the technology to point of characterised by flow cytometric analysis to determine the care applications. distribution of PBMCs and erythrocytes. To distinguish between nucleated PBMCs and erythrocytes, cells were Author Statement stained with Höchst 33342 (compare figure 4). The diagrams The authors would like to express great appreciation for the financial support to the German Federal Ministry of Education and Research (BMBF) for the project “SIMPLE-IVD” within the “Innovations for Tomorrow’s Production, Services, and Work” program (FK 02P18C100) implemented by the Project Management Agency Karlsruhe (PTKA). References Figure 4: Scatter plots of single cell signals of Höchst 33342 [1] Kaufmann SHE. Basiswissen Immunologie. Berlin: Springer (H33342) vs. foreward scatter(FSC): A) Buffy coat; A) Purified sample [2] Ali SA, Shi V, Maric I, et al. T cells expressing an anti-B-cell maturation antigen chimeric antigen receptor cause show the gated populations in red, green and violet remissions of multiple myeloma. Blood 2016; 128: 1688– representing the lymphocytes, granulocytes and monocytes. The black gate shows the number of erythrocytes. Compared [3] Grützkau A, Radbruch A. Small but mighty: how the MACS- to the buffy coat the distribution of cell populations after technology based on nanosized superparamagnetic purification showed a significant increase in the proportion of particles has helped to analyze the immune system within lymphocytes in the sample with a mean of 80,1% (compare the last 20 years. Cytometry. Part A : the journal of the table 1) and a maximum value of 93.2 %. Thus, the selectivity International Society for Analytical Cytology 2010; 77: 643– of the microfluidic cartridge is comparable to the established TACS cartridge (selectivity > 90%) [9], which was the initial [4] Weiss R, Gerdes W, Leonhardt F, Berthold R, Sack U, benchmark. Nevertheless, the repeatability of the experiments Grahnert A. A comparative study of two separation methods has to be further improved by geometric changes of the to isolate monocytes. Cytometry. Part A : the journal of the cartridge outlet to minimize erythrocyte accumulation during International Society for Analytical Cytology 2019; 95: 234– the rinsing of the negative fraction. [5] Laurien E, Oertel H. Numerische Strömungsmechanik. 5th Table 1: Mean amount of selected cells from three microfluidic ed. Wiesbaden: Springer Vieweg 2013. cartridge tests: L – lymphocytes; E – erythrocytes; M – monocytes; [6] Sonntag F, Grünzner S, Schmieder F, Busek M, Klotzbach G - granulocytes U, Franke V. Multilayer based lab-on-a-chip-systems for L [%] E [%] M [%] G [%] substance testing. In: Klotzbach U, Washio K, Arnold CB, Buffy coat 38,7 55,4 2,5 3,6 editors. Laser-based Micro- and Nanoprocessing IX: SPIE; Microfluidic 2015. 93510C (SPIE Proceedings). 80,1 ± 10,01 19,7 ± 10,0 0,0 ± 0,01 0,1 ± 0,1 cartridge [7] Busek M. Modellierung und Regelung der Nährstoffversorgung in mikrofluidischen Zellkultursystemen. Dresden: TUDpress 2018. 5 Conclusion [8] Sung JH, Wang Y, Shuler ML. Strategies for using mathematical modeling approaches to design and interpret This work shows that simulation based down scaling is a multi-organ microphysiological systems (MPS). APL suitable method to transfer a medium volume cartridge for bioengineering 2019; 3: 21501. blood cell separation from millilitre to microliter scale. The [9] IBA GmbH. Isolation Kit forCD3+ cellsfrom Whole Blood prototype of the microfluidic cartridge yields to similar purity (human)or Buffy Coat (human) with FABian®. in the isolation of lymphocytes compared to the established

Journal

Current Directions in Biomedical Engineeringde Gruyter

Published: Sep 1, 2020

Keywords: TACS; microfluidic; in-vitro diagnostics; simulation; Cell separation

There are no references for this article.