Immunotrafficking Lab

Physics and Astronomy, University of Manitoba

 

Overview of Research

Cell migration is critically underlying many physiological processes ranging from immune responses to tissue development and to cancer metastasis. The cell migration mechanism itself is complex. Furthermore, the diverse environmental guiding factors and their spatiotemporal interactions with cells enable highly controlled directional cell migration and trafficking in tissues. Our work aims to dissect these mechanisms with a focus on chemical and electrical guidance. Using microfluidic devices, we quantitatively measure and analyze the migration of different cell types in defined physiologically-relevant chemical gradients and electric fields. Furthermore, we develop quantitative models to investigate the cellular mechanisms for cell chemotactic and electrotactic navigation. On the application side, we are developing microfluidic point-of-care systems for disease biomarker monitoring and new easy-to-use microfluidic tools for life science research.

 

 

Trends in Cell Biology, 2011, Vol. 21, 489-497.
 

 

 

MS2 Technology, K. Yang et al, Lab on Chip, 2016

 

[1] K. Yang, J.D. Wu, S. Santos, Y. Liu, L. Zhu and F. Lin, “Recent development of portable imaging platforms for cell-based assays”, Biosensors and Bioelectronics, 2019, 124-125, 150-160.
[2] Z. Rovei Miab, F. Lin and J. Anderson, “Emerging development of microfluidic-based approaches to improve studies of muscle cell migration”, Tissue Engineering, Part B, 2018; doi: 10.1089/ten.TEB.2018.0181.
[3] J.D. Wu, M.L. Dong, C. Rigatto, Y. Liu and F. Lin, “Lab-on-chip technology for chronic disease diagnosis”, npj Digital Medicine, 2018, 1:7; doi: 10.1038/s41746-017-0014-0.

[4]
J.D. Wu, M.L. Dong, S. Santos, C. Rigatto, Y. Liu and F. Lin, “Lab-on-chip systems for detection of cardiovascular disease and cancer biomarkers”, Sensors, 2017, 17(12), 2934.

[5] X.O. Ren, D. Levin and F. Lin, “Cell migration research based on organ-on-chip related approaches”, Special Issue of “Integrated Microfluidics for Chemical Synthesis and Analysis”, Micromachines, 2017, 8(11), 324.
Featured as the cover image
[6] K. Yang, H. Peretz-Soroka, Y. Liu and F. Lin, “Novel Developments of Mobile Sensing Based on the Integration of Microfluidic Devices and Smartphone”, Lab on a Chip, 2016, 16, 943-958. Featured as the back cover image
[7]
J.D. Wu and F. Lin, Chapter 7, “Micro-engineered tools for studying cell migration in electric fields”, for the book ‘Integrative Mechanobiology: Micro and Nano Techniques in Cell Mechanobiology’, Y. Sun, D.H. Kim, and C. Simmons (Eds.), Cambridge University Press, 2015.
[8] J. Li and F. Lin, "Microfluidic Devices for Studying Chemotaxis and Electrotaxis", Trends in Cell Biology, 2011, Vol. 21, 489-497.

[9] J.D. Wu and F. Lin, “Recent Developments in Electrotaxis Assays". Advances in Wound Care. 2013, doi:10.1089/wound.20.

[10] J.D. Wu, X. Wu and F. Lin, “Recent Developments in Microfluidics-Based Chemotaxis Studies". Lab on Chip. 2013, 13(13):2484-99.

[11] F. Lin and C. Pullar, "Inflammatory Cell Electrotaxis", Chapter 8, The Physiology of Bioelectricity in Development, Tissue Regeneration and Cancer, CRC Press, 2011.

[12] F. Lin, “A Microfluidics-Based Method for Analyzing Leukocyte Migration to Chemoattractant Gradients”, Methods in Enzymology, 2009, Vol. 461, Chapter 15, 333-347.

[13] F. Lin, B.G. Chung, W. Saadi, and N.L. Jeon, “Gradient-Generating Microfluidic Devices for Cell Biology Research”, Chapter 2, Micro- and Nano-Engineering of the Cell Microenvironment: Technologies and Applications, Artech House Publishing Inc, March 2008.

 

Immune cell chemotaxis
Immune cells are required to effectively navigate through complex chemoattractant fields in tissues and traffic to the target site to perform their immune functions. Current research focuses on investigating leukocyte migration and chemotaxis in physiologically-relevant co-existing chemoattractant fields. Using microfluidics-based experimental approach, cell biology and immunology methods, as well as mathematical modeling, we are studying the chemical guiding mechanisms underlying lymphocyte migration and trafficking in secondary lymphoid tissues. In collaboration with industrial partners, we have also studied the effect of bacteria-produced toxins on lymphocyte migration using microfluidic devices toward better understanding the mechanisms for bacteria induced intestinal diseases and thus facilitating the development of therapeutic applications.

 

 

Combinatorial guidance model

 

 

 

Human blood T cell chemotaxis in a microfluidic device

 

 

 

Jurkat CCR7-EGFP transfectants chemotaxis in a microfluidic device

The dual-docking chip (D2-Chip) for studying neutrophil chemotaxis and the memtory effect



Machine learning for cell migration data analysis

 

[1] J.D. Wu, A. Kumar-Kanojia, S. Hombach-Klonisch, T. Klonisch and F. Lin, "A radial microfluidic platform for higher throughput chemotaxis studies with individual gradient control", Lab on a Chip, 2018,18, 3855 - 3864.
[2]
K. Yang, H. Peretz-Soroka, J.D. Wu, L. Zhu, X.L. Cui, M. Zhang, C. Rigatto, Y. Liu and F. Lin, "Fibroblast growth factor 23 weakens chemotaxis of human blood neutrophils in microfluidic devices", Scientific Reports, 2017, 7: 3100, DOI: 10.1038/s41598-017-03210-0.

[3]. K. Yang, J.D. Wu, G.Q. Xu, D.X. Xie, H. Peretz-Soroka, S. Santos, M. Alexander, L. Zhu, M. Zhang, Y. Liu and F. Lin, “A dual-docking microfluidic cell migration assay (D2-Chip) for testing neutrophil chemotaxis and the memory effect", Integrative Biology, 2017, 9, 303 - 312. Featured as the back cover image.
[4]
D.X. Xie, Z.H. Liu, J.D. Wu, W.F. Feng, K. Yang, J.X. Deng, G.H. Tian, S. Santos, X.L. Cui and F. Lin, "The effects of activin A on the migration of human breast cancer cells and neutrophils and their migratory interaction", Experimental Cell Research, 2017, http://dx.doi.org/10.1016/j.yexcr.2017.05.003.
[5]
I.
Halilovic, J.D. Wu, M. Alexander, F. Lin, “Neutrophil Migration under Spatially-Varying Chemoattractant Gradient Profiles”, Biomedical Microdevices, 2015, Jun;17(3):9963.

[6] Xun Wu, Jiandong Wu, Hongzhao Li, Daniel F. Legler, Aaron J. Marshall, Francis Lin, “Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator”, Journal of Immunological Methods, 419 (2015) 9–17.
[7] D. Wu, A.G. Joyee, S. Nandagopal, M. Lopez, X.L. Ma, J. Berry and F. Lin, "Effects of C. difficile toxin A and B on human T lymphocyte migration".
Toxins, 2013. 5 (5), 926-938.

[8] D. Wu and F. Lin, "Modeling Cell Gradient Sensing and Migration in Competing Chemoattractant Gradients", PLoS ONE, 2011, 6(4): e18805.

[9] S. Nandagopal, D. Wu and F. Lin, "Combinatorial Guidance by CCR7 Ligands for T Lymphocyte Migration in Co-Existing Chemokine Fields", PLoS ONE, 2011, 6(3): e18183.

[10] F. Lin and E.C. Butcher, “Modeling the Role of Homologous Receptor Desensitization in Cell Gradient Sensing”, Journal of Immunology, 2008, 181(12): 8335-43.

[11] F. Lin and E.C. Butcher, “T Cell Chemotaxis in a Simple Microfluidic Device”, Lab on a Chip, 2006, 6: 1462-1469

[12] F. Lin, C. M-C Nguyen, S.J. Wang, W. Saadi, S.P. Gross and N.L. Jeon, “Neutrophil Migration in Opposing Chemoattractant Gradients Using Microfluidic Chemotaxis Devices”, Annals of Biomedical Engineering, 2005, 33: 473-480.

 

Electrotaxis
Physiological electric fields widely exist in tissues such as healing wound, to which immune cells are exposed. We have previously demonstrated electric field directed lymphocyte migration (electrotaxis) in vitro and in vivo. Such electrical guidance may mediate immune responses and may be used for therapeutic applications. Current studies continue to quantitatively analyze immune cell electrotaxis and to explore its underlying mechanisms. Furthermore, we are developing microfluidics-based approach to study the interactions of chemical and electrical cues for directing immune cell migration. In addition, we have extended the electrotaxis study to other cell types such as breast cancer cells, adult stem cells and epithelial cells. In particular, in collaboration with the Zhao lab at UC Davis, we have been studying collective epithelial cell migration and electrotaxis in wound healing by combining experimental and modeling approaches.

 

 

High-throughput screening of white blood cell electrotaxis in vitro

 

Lymphocyte electrotaxis in the ear tissue of a trangenetic mouse imaged by intravital confocal microscopy

Lymphocyte electrotaxis in vitro and in vivo

J. Immuno., 2008, 181(4): 2465-71. Copyright 2008, AAI Inc.


Experimental and modeling studies of collective cell migration in wound healing

 

Bioenergetic model for amoeboid-like cell motility and electrotaxis

 

[1] H. Peretz-Soroka, R. Tirosh, J. Hipolito, E. Huebner, M. Alexander, J. Fiege, F. Lin, “A bioenergetic mechanism for amoeboid-like cell motility profiles tested in a microfluidic electrotaxis assay”, Integrative Biology, 2017, 9, 844 - 856. Featured as the front cover image.
[2]
Y. Zhang, G.Q. Xu, R.M Lee, Z.J. Zhu, J.D. Wu, S. Liao, G. Zhang, Y.H. Sun, A. Mogilner, W. Losert, T.R. Pan, F. Lin, Z.P. Xu, M. Zhao, “Collective Cell Migration has Distinct Directionality and Speed Dynamics”, Cellular and Molecular Life Sciences, 2017, 74(20), 3841-3850.

[3] D. Wu, X.L. Ma and F. Lin, "DC Electric Fields Direct Breast Cancer Cell Migration, Induce EGFR Polarization and Increase the Intracellular Level of Calcium Ion". Cell Biochemistry and Biophysics, 2013, 67 (3):1115-1125.
[4] L. Li, R. Hartley, B. Reiss, Y.H. Sun, J. Pu, D. Wu, F. Lin, T. Hoang, S. Yamada, J.X. Jiang, M. Zhao, "E-cadherin plays an essential role in collective directional migration of large epithelial sheets", Cellular and Molecular Life Sciences, 2012, 69(16):2779-89.

[5] J. Li, L. Zhu, M. Zhang and F. Lin, "Microfluidic Device for Studying Cell Migration in Single or Co-Existing Chemical Gradients and Electric Fields", Biomicrofluidics, 2012, 6, 024121.

[6] D. Wu and F. Lin, "A Receptor Electromigration-Based Model for Cellular Electrotactic Sensing and Migration", Biochem Biophys Res Commun, 2011, 411, 695-701.

[7] J. Li, S. Nandagopal, D. Wu, S.F. Romanuik, K. Paul, D.J. Thomson and F. Lin, "Activated T Lymphocytes Migrate Toward the Cathode of DC Electric Fields in Microfluidic Devices", Lab on a Chip, 2011, 11(7), 1298 -1304.

[8] F. Lin, F. Baldessari, C.C. Gyenge, T. Sato, R.D. Chambers, J.G. Santiago and E.C. Butcher, “Lymphocyte Electrotaxis in vitro and in vivo”, Journal of Immunology, 2008, 181(4): 2465-71.

 

Stem cell migration
Stem cells offer important options for regenerative medicine owing to its self-renewal and differentiation ability. Adipose-derived stem cells (ASC) are promising stem cell source for treating various degenerative diseases. However, the effectiveness of ASC therapy is limited by its low homing efficiency to the target tissues and low purity. In collaboration with the Tian group at the NRC, we are using microfluidic devices to study ASC migration and to develop new ASC selection methods.

 


 

On-chip selection of chemotactic stem cells


[1] N. Wadhawan, H. Kalkat, K. Natarajan, X.L. Ma, S. Gajjeraman, S. Nandagopal, N. Hao, J. Li, M. Zhang, J.X. Deng, B. Xiang, S. Mzengeza, D. Freed, R. Arora, G.H. Tian and F. Lin, "Growth and Positioning of Adipose-Derived Stem Cells in Microfluidic Devices". Lab on a Chip, 2012, 12 (22), 4829 - 4834.

[2] K. Natarajan, C. Tian, B. Xiang, C. Chi, J.X. Deng, R.D. Zhang, D.H. Freed, R.C. Arora, G.H. Tian, F. Lin, “Selection of chemotactic adipose-derived stem cells using a microfluidic gradient generator”, RSC Advances, 2015, 5, 6332-6339.

 

 

Microfluidics
Microfluidics is a fast growing emerging research field. Its development is changing the way life science research is performed and is enabling a broad range of medical applications. We are developing new microfluidic devices for cell migration studies. The focused applications are on chemical gradient generation for chemotaxis studies and controlled electric field for electrotaxis devices, and strategies for studying cell migration in superimposed chemical gradients and electric fields.

 


Microfluidic device for generating competing chemical gradient and electric field

Biomicrofluidics, 2012, 6, 024121.

 

 




All-on-chip chemotaxis test

Technology, 2016



High-throughput microfluidic cell migration assay (D3-Chip)

Scientific Reports, 2017

 

[1] J.D. Wu, A. Kumar-Kanojia, S. Hombach-Klonisch, T. Klonisch and F. Lin, "A radial microfluidic platform for higher throughput chemotaxis studies with individual gradient control", Lab on a Chip, 2018, doi: 10.1039/C8LC00981C.
[2] K. Yang, H. Peretz-Soroka, J.D. Wu, L. Zhu, X.L. Cui, M. Zhang, C. Rigatto, Y. Liu and F. Lin, "Fibroblast growth factor 23 weakens chemotaxis of human blood neutrophils in microfluidic devices", Scientific Reports, 2017, 7: 3100, DOI: 10.1038/s41598-017-03210-0.

[3] J.D. Wu, C. Hillier, P. Komenda, R. Lobato de Faria, S. Santos, D. Levin, M. Zhang and F. Lin, “An all-on-chip method for testing neutrophil chemotaxis induced by fMLP and COPD patient’s sputum”, Technology, 2016, 4(2), 104-9.

[4] J. Li, L. Zhu, M. Zhang and F. Lin, "Microfluidic Device for Studying Cell Migration in Single or Co-Existing Chemical Gradients and Electric Fields", Biomicrofluidics, 2012, 6, 024121.

[5] J. Li, S. Nandagopal, D. Wu, S.F. Romanuik, K. Paul, D.J. Thomson and F. Lin, "Activated T Lymphocytes Migrate Toward the Cathode of DC Electric Fields in Microfluidic Devices", Lab on a Chip, 2011, 11(7), 1298 -1304.
[6] Francis Lin and Eugene C. Butcher, “T Cell Chemotaxis in a Simple Microfluidic Device”, Lab on a Chip, 2006, 6: 1462-1469.
[7] Bong Geun Chung, Francis Lin (co-first author) and Noo Li Jeon, “A Microfluidic Multi-Injector for Gradient Generation”, Lab on a Chip, 2006, 6(6):764-8.

[8] Francis Lin, Wajeeh Saadi, Seog Woo Rhee, Shur-Jen Wang, Sukant Mittal* and Noo Li Jeon, “Generation of Dynamic Temporal and Spatial Concentration Gradients Using Microfluidic Devices”, Lab on a Chip, 2004, 4: 164-167.


Portable microfluidic systems and point-of-care applications
Miniaturization, low costs, high-throughput and integration are important features of microfluidic devices, enabling new applications for both basic research and clinical studies. To remove the barrier of accepting microfluidic systems as a common research tool in life science labs, we are developing integrated, low cost and portable microfluidic systems for live cell analysis. Furthermore, through collaborations with clinical researchers and international partnerships, we are interested in developing and testing integrated lab-on-chip systems suitable for point-of-care disease biomarker monitoring.





Neutrophil chemotaxis to a COPD sputum gradient


Smartphone-based cell migration assay

Rapid and low-cost chronic disease protein biomarker measurement

 

[1] J.D. Wu, D. Tomsa, M. Zhang, P. Komenda, N. Tangri, C. Rigatto and F. Lin, "A Passive Mixing Microfluidic Urinary Albumin Chip (UAL-Chip) for Chronic Kidney Disease Assessment", ACS Sensors, 2018, 3(10):2191-2197.
[2] K. Yang, J.D. Wu, H. Peretz-Soroka, L. Zhu, Z.G. Li, Y.S. Sang, J. Hipolito, M. Zhang, S. Santos, C. Hillier, R. Lobato de Faria, Y. Liu, F. Lin, “Mkit: A Cell Migration Assay Based on Microfluidic Device and Smartphone”, Biosensors and Bioelectronics, 2018, 99:259-267.
[3] K. Yang, J.D. Wu, L. Zhu, Y. Liu, M. Zhang and F. Lin, “An all-on-chip method for rapid neutrophil chemotaxis analysis directly from a drop of blood”, J. Vis. Exp., (124), e55615, doi:10.3791/55615 (2017). http://www.jove.com/video/55615
[4].
M.L. Dong, J.D. Wu, Z.M. Ma, H. Peretz-Soroka, M. Zhang, P. Komenda, N. Tangri, Y. Liu, C. Rigatto and F. Lin, "
Rapid and low-cost CRP measurement by integrating a paper-based microfluidic immunoassay with smartphone (CRP-Chip)", Special issue of "State-of-the-Art Sensors Technology in Canada 2017", Sensors, 2017, 17(4), 684.
[5]. F. Lin and J.D. Wu, “Microfluidic chemotaxis assay”, US provisional patent, filed on Apr. 8, 2016, Application number: US 62/312,570.
[6]
J.D. Wu, C. Hillier, P. Komenda, R. Lobato de Faria, S. Santos, D. Levin, M. Zhang and F. Lin, “An all-on-chip method for testing neutrophil chemotaxis induced by fMLP and COPD patient’s sputum”, Technology, 2016, doi:10.1142/S2339547816500035.
[7] K. Yang, H. Peretz-Soroka, Y. Liu and F. Lin, “Novel Developments of Mobile Sensing Based on the Integration of Microfluidic Devices and Smartphone”, Lab on a Chip, 2016, 16, 943-958. Featured as the back cover image
[8] J.D. Wu, L.P. Ouyang, N. Wadhawana, J. Li, M. Zhang, S. Liao, D. Levin and F. Lin, "A compact microfluidic system for cell migration studies", Biomedical Microdevices, 2014, 16(4): 521-528.

[9]. J.D. Wu, C. Hillier, P. Komenda, R. Lobato de Faria, D. Levin, M. Zhang and F. Lin, “A microfluidic platform for evaluating neutrophil chemotaxis induced by sputum from COPD patients”, PLoS ONE, 2015 May 11;10(5):e0126523.

[10] J.D. Wu, L.P. Ouyang, M. Zhang, S. Liao, C. Hillier, P. Komenda, R.L. de Faria and F. Lin, “Assessing neutrophil chemotaxis in COPD using a compact microfluidic system”, The 36th Annual International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC’14), Chicago, U.S.A., August 26-30, 2014.
[11] F. Lin and J.D. Wu, “Low-Cost Portable Microfluidic System for Cell Migration Studies”, PCT application, Filed on Sept. 11, 2013, Application No: PCT/CA2013/050697.

[12] L. Zhu, L. Li, D. An, M. Wang, L. Zhang, Y.K. Wang, Z. Li, Y. Liu, G. Zhang, F. Lin, “An Integrated Microfluidic Real-Time PCR System for Pathogen Detection”, TechConnect World 2012, Santa Clara, California, June 18-21, 2012.