Home LiteratureArticle Details
PMID: 18094760 Published · ppublish English Journal Article Review

Biomolecular gradients in cell culture systems.

Lab on a chip ·Vol. 8 ·No. 1 ·2008-01-00 ·Pages 34-57

Keenan TM, Folch A

Abstract

Biomolecule gradients have been shown to play roles in a wide range of biological processes including development, inflammation, wound healing, and cancer metastasis. Elucidation of these phenomena requires the ability to expose cells to biomolecule gradients that are quantifiable, controllable, and mimic those that are present in vivo. Here we review the major biological phenomena in which biomolecule gradients are employed, traditional in vitro gradient-generating methods developed over the past 50 years, and new microfluidic devices for generating gradients. Microfluidic gradient generators offer greater levels of precision, quantitation, and spatiotemporal gradient control than traditional methods, and may greatly enhance our understanding of many biological phenomena. For each method, we outline the salient features, capabilities, and applications.

MeSH Terms
Animals Biological Factors/metabolism,pharmacology Cell Culture Techniques/instrumentation,methods Cell Physiological Phenomena/drug effects Microfluidic Analytical Techniques/methods Substrate Specificity Time Factors
Chemicals
Biological Factors
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Keenan Thomas M
Department of Bioengineering, University of Washington, Washington 98195, USA.
Folch Albert
References (111)
111 references, click to expand
  1. A sensitive, versatile microfluidic assay for bacterial chemotaxis.
    Proc Natl Acad Sci U S A. 2003 Apr 29;100(9):5449-54 PMID: 12704234
  2. Morphological changes and membrane potential of human granulocytes under influence of chemotactic peptide and/or echo-virus, type 9.
    Klin Wochenschr. 1988 May 16;66(10):434-6 PMID: 3398429
  3. Concentration and separation of proteins in microfluidic channels on the basis of transverse IEF.
    Anal Chem. 2001 Apr 1;73(7):1627-33 PMID: 11321320
  4. Adaptation in the chemotactic guidance of nerve growth cones.
    Nature. 2002 May 23;417(6887):411-8 PMID: 11986620
  5. Chemotaxis of capacitated rabbit spermatozoa to follicular fluid revealed by a novel directionality-based assay.
    Biol Reprod. 2002 Nov;67(5):1565-71 PMID: 12390889
  6. Increased velocity and induction of chemotactic response in mouse spermatozoa by follicular and oviductal fluids.
    J Reprod Fertil. 1999 Jan;115(1):23-7 PMID: 10341719
  7. Chemoattractant receptor cross talk as a regulatory mechanism in leukocyte adhesion and migration.
    Eur J Immunol. 1997 Oct;27(10):2571-8 PMID: 9368612
  8. A parallel-gradient microfluidic chamber for quantitative analysis of breast cancer cell chemotaxis.
    Biomed Microdevices. 2006 Jun;8(2):109-18 PMID: 16688570
  9. Chemotaxis assays of mouse sperm on microfluidic devices.
    Anal Chem. 2006 May 15;78(10):3354-9 PMID: 16689537
  10. Signal transduction underlying growth cone guidance by diffusible factors.
    Curr Opin Neurobiol. 1999 Jun;9(3):355-63 PMID: 10395576
  11. Effects of synthetic micro- and nano-structured surfaces on cell behavior.
    Biomaterials. 1999 Mar;20(6):573-88 PMID: 10213360
  12. Regiospecific control of protein expression in cells cultured on two-component counter gradients of extracellular matrix proteins.
    Langmuir. 2005 Mar 29;21(7):3061-8 PMID: 15779985
  13. The interpretation of morphogen gradients.
    Development. 2006 Feb;133(3):385-94 PMID: 16410409
  14. Continuous concentration of bacteria in a microfluidic flow cell using electrokinetic techniques.
    Electrophoresis. 2001 Jan;22(2):355-62 PMID: 11288905
  15. Microcontact printing of axon guidance molecules for generation of graded patterns.
    Nat Protoc. 2006;1(3):1322-8 PMID: 17406418
  16. Integrating conflicting chemotactic signals. The role of memory in leukocyte navigation.
    J Cell Biol. 1999 Nov 1;147(3):577-88 PMID: 10545501
  17. Effects of flow and diffusion on chemotaxis studies in a microfabricated gradient generator.
    Lab Chip. 2005 Jun;5(6):611-8 PMID: 15915253
  18. Recent advances in microcontact printing.
    Anal Bioanal Chem. 2005 Feb;381(3):591-600 PMID: 15696278
  19. Quantitative analysis of molecular interaction in a microfluidic channel: the T-sensor.
    Anal Chem. 1999 Dec 1;71(23):5340-7 PMID: 10596213
  20. Neuronal chemotaxis: chick dorsal-root axons turn toward high concentrations of nerve growth factor.
    Science. 1979 Nov 30;206(4422):1079-80 PMID: 493992
  21. Microengineering of cellular interactions.
    Annu Rev Biomed Eng. 2000;2:227-56 PMID: 11701512
  22. Targeting key steps in metastatic tumour progression.
    Curr Opin Genet Dev. 2005 Feb;15(1):77-86 PMID: 15661537
  23. Characterization of a membrane-based gradient generator for use in cell-signaling studies.
    Lab Chip. 2006 Mar;6(3):389-93 PMID: 16511622
  24. Surface engineering approaches to micropattern surfaces for cell-based assays.
    Biomaterials. 2006 Jun;27(16):3044-63 PMID: 16458351
  25. A three-channel microfluidic device for generating static linear gradients and its application to the quantitative analysis of bacterial chemotaxis.
    Lab Chip. 2006 Mar;6(3):381-8 PMID: 16511621
  26. The role of the floor plate in axon guidance.
    Annu Rev Neurosci. 1995;18:497-529 PMID: 7605072
  27. A rapid diffusion immunoassay in a T-sensor.
    Nat Biotechnol. 2001 May;19(5):461-5 PMID: 11329017
  28. Differential effects of EGF gradient profiles on MDA-MB-231 breast cancer cell chemotaxis.
    Exp Cell Res. 2004 Oct 15;300(1):180-9 PMID: 15383325
  29. Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device.
    Nat Biotechnol. 2002 Aug;20(8):826-30 PMID: 12091913
  30. Multistep navigation and the combinatorial control of leukocyte chemotaxis.
    J Cell Biol. 1997 Dec 1;139(5):1349-60 PMID: 9382879
  31. Generation of complex, static solution gradients in microfluidic channels.
    J Am Chem Soc. 2006 Apr 5;128(13):4194-5 PMID: 16568971
  32. Fabrication of gradient hydrogels using a microfluidics/photopolymerization process.
    Langmuir. 2004 Jun 22;20(13):5153-6 PMID: 15986641
  33. Gradient steepness influences the pathfinding decisions of neuronal growth cones in vivo.
    J Neurosci. 2003 Jan 1;23(1):193-202 PMID: 12514216
  34. Analysis of the directed and nondirected movement of human granulocytes: influence of temperature and ECHO 9 virus on N-formylmethionylleucylphenylalanine-induced chemokinesis and chemotaxis.
    J Cell Biol. 1983 Jun;96(6):1708-16 PMID: 6853601
  35. The molecular genetics of chemotaxis: sensing and responding to chemoattractant gradients.
    Bioessays. 2000 Jul;22(7):603-15 PMID: 10878573
  36. Dose-dependent priming or desensitization induced by chemotactic agents in chemiluminescence experiments with canine and human neutrophils.
    Vet Immunol Immunopathol. 1998 Nov 6;66(1):11-24 PMID: 9847017
  37. Growth-cone attraction to netrin-1 is converted to repulsion by laminin-1.
    Nature. 1999 Sep 2;401(6748):69-73 PMID: 10485706
  38. Measurement of cell migration in response to an evolving radial chemokine gradient triggered by a microvalve.
    Lab Chip. 2006 Jul;6(7):849-56 PMID: 16804588
  39. Generation of stable complex gradients across two-dimensional surfaces and three-dimensional gels.
    Langmuir. 2007 Oct 23;23(22):10910-2 PMID: 17910490
  40. Conversion of neuronal growth cone responses from repulsion to attraction by cyclic nucleotides.
    Science. 1998 Sep 4;281(5382):1515-8 PMID: 9727979
  41. Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors.
    J Cell Biol. 1977 Nov;75(2 Pt 1):606-16 PMID: 264125
  42. Generation of linear and non-linear concentration gradients along microfluidic channel by microtunnel controlled stepwise addition of sample solution.
    Lab Chip. 2007 Oct;7(10):1371-3 PMID: 17896024
  43. Analyzing chemotaxis using the Dunn direct-viewing chamber.
    Methods Mol Biol. 1997;75:449-57 PMID: 9276291
  44. A comparative study of the neutrophil stimulatory activity in vitro and pro-inflammatory properties in vivo of 72 amino acid and 77 amino acid IL-8.
    J Immunol. 1992 Jan 1;148(1):106-11 PMID: 1727857
  45. Role of interleukin-8 in neutrophil signaling.
    Curr Opin Hematol. 2000 May;7(3):178-82 PMID: 10786656
  46. Cell lysis and protein extraction in a microfluidic device with detection by a fluorogenic enzyme assay.
    Anal Chem. 2002 Apr 15;74(8):1798-804 PMID: 11985310
  47. Cancer and the chemokine network.
    Nat Rev Cancer. 2004 Jul;4(7):540-50 PMID: 15229479
  48. The ionic basis of chemotaxis. Separate cation requirements for neutrophil orientation and locomotion in a gradient of chemotactic peptide.
    Am J Pathol. 1980 Mar;98(3):749-68 PMID: 6767408
  49. Effective neutrophil chemotaxis is strongly influenced by mean IL-8 concentration.
    Biochem Biophys Res Commun. 2004 Jun 25;319(2):576-81 PMID: 15178445
  50. A new chemotaxis assay shows the extreme sensitivity of axons to molecular gradients.
    Nat Neurosci. 2004 Jun;7(6):678-82 PMID: 15162167
  51. The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes.
    J Exp Med. 1962 Mar 1;115:453-66 PMID: 13872176
  52. Mouse spermatozoa modify their motility parameters and chemotactic response to factors from the oocyte microenvironment.
    Int J Androl. 1998 Aug;21(4):201-6 PMID: 9749350
  53. Propagation of activity-dependent synaptic depression in simple neural networks.
    Nature. 1997 Jul 31;388(6641):439-48 PMID: 9242402
  54. Human neural stem cell growth and differentiation in a gradient-generating microfluidic device.
    Lab Chip. 2005 Apr;5(4):401-6 PMID: 15791337
  55. Cell migration and polarity on microfabricated gradients of extracellular matrix proteins.
    Langmuir. 2006 Apr 25;22(9):4250-8 PMID: 16618172
  56. Universal microfluidic gradient generator.
    Anal Chem. 2006 May 15;78(10):3472-7 PMID: 16689552
  57. Echo 9 virus-induced order-disorder transition of chemotactic response of human polymorphonuclear leucocytes: phenomenology and molecular biology.
    Blood Cells. 1984;10(1):79-106 PMID: 6487817
  58. Basic fibroblast growth factor (Fgf2) is necessary for cell proliferation and neurogenesis in the developing cerebral cortex.
    J Neurosci. 2000 Jul 1;20(13):5012-23 PMID: 10864959
  59. A hydrogel-based microfluidic device for the studies of directed cell migration.
    Lab Chip. 2007 Jun;7(6):763-9 PMID: 17538719
  60. Essential role of filopodia in chemotropic turning of nerve growth cone induced by a glutamate gradient.
    J Neurosci. 1996 Feb 1;16(3):1140-9 PMID: 8558243
  61. Turning of nerve growth cones induced by neurotransmitters.
    Nature. 1994 Mar 10;368(6467):140-4 PMID: 8139655
  62. Neutrophil migration in opposing chemoattractant gradients using microfluidic chemotaxis devices.
    Ann Biomed Eng. 2005 Apr;33(4):475-82 PMID: 15909653
  63. Chemotactic desensitization of neutrophils demonstrates interleukin-8 (IL-8)-dependent and IL-8-independent mechanisms of transmigration through cytokine-activated endothelium.
    Immunology. 1993 Mar;78(3):491-7 PMID: 8478031
  64. Cells on chips.
    Nature. 2006 Jul 27;442(7101):403-11 PMID: 16871208
  65. Chemotaxis and chemokinesis of malignant mesothelioma cells to multiple growth factors.
    Anticancer Res. 2004 May-Jun;24(3a):1625-30 PMID: 15274332
  66. The effects of radiographic contrast media on leucocyte orientation.
    Br J Radiol. 1993 Sep;66(789):778-80 PMID: 8220946
  67. Gradients of substrate-bound laminin orient axonal specification of neurons.
    Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12542-7 PMID: 12237407
  68. Application of inkjet printing to tissue engineering.
    Biotechnol J. 2006 Sep;1(9):910-7 PMID: 16941443
  69. Hierarchical organization of guidance receptors: silencing of netrin attraction by slit through a Robo/DCC receptor complex.
    Science. 2001 Mar 9;291(5510):1928-38 PMID: 11239147
  70. Electrical activity modulates growth cone guidance by diffusible factors.
    Neuron. 2001 Feb;29(2):441-52 PMID: 11239434
  71. Netrin-1 is required for commissural axon guidance in the developing vertebrate nervous system.
    Cell. 1996 Dec 13;87(6):1001-14 PMID: 8978605
  72. Polarization of chemoattractant receptor signaling during neutrophil chemotaxis.
    Science. 2000 Feb 11;287(5455):1037-40 PMID: 10669415
  73. Conserved roles for Slit and Robo proteins in midline commissural axon guidance.
    Neuron. 2004 Apr 22;42(2):213-23 PMID: 15091338
  74. cAMP-induced switching in turning direction of nerve growth cones.
    Nature. 1997 Jul 17;388(6639):275-9 PMID: 9230436
  75. Biology on a chip: microfabrication for studying the behavior of cultured cells.
    Crit Rev Biomed Eng. 2003;31(5-6):423-88 PMID: 15139302
  76. EphB receptors and ephrin-B3 regulate axon guidance at the ventral midline of the embryonic mouse spinal cord.
    J Neurosci. 2006 Aug 30;26(35):8909-14 PMID: 16943546
  77. Dynamics of a chemoattractant receptor in living neutrophils during chemotaxis.
    Mol Biol Cell. 1999 Apr;10(4):1163-78 PMID: 10198064
  78. Glycolipid storage material in Fabry's disease: a study by electron microscopy, freeze-fracture, and digital image analysis.
    J Struct Biol. 1990 Mar;103(1):40-7 PMID: 2118788
  79. Polymorphonuclear leukocyte chemotaxis induced by zinc, copper and nickel in vitro.
    Biochim Biophys Acta. 1995 Oct 19;1245(2):145-52 PMID: 7492570
  80. Asymmetric modulation of cytosolic cAMP activity induces growth cone turning.
    J Neurosci. 1992 Apr;12(4):1253-61 PMID: 1372932
  81. Growth cone navigation in substrate-bound ephrin gradients.
    Development. 2006 Jul;133(13):2487-95 PMID: 16763203
  82. MAPK-mediated bimodal gene expression and adaptive gradient sensing in yeast.
    Nature. 2007 Mar 1;446(7131):46-51 PMID: 17310144
  83. Pituitary development: regulatory codes in mammalian organogenesis.
    Science. 2002 Mar 22;295(5563):2231-5 PMID: 11910101
  84. Neutrophil polarization: spatiotemporal dynamics of RhoA activity support a self-organizing mechanism.
    Proc Natl Acad Sci U S A. 2006 Mar 7;103(10):3639-44 PMID: 16537448
  85. Chemotactically-induced redistribution of CD43 as related to polarity and locomotion of human polymorphonuclear leucocytes.
    Biol Cell. 2003 Jul;95(5):265-73 PMID: 12941524
  86. Signaling mechanisms for regulation of chemotaxis.
    Cell Res. 2005 Jan;15(1):52-6 PMID: 15686628
  87. cAMP-dependent growth cone guidance by netrin-1.
    Neuron. 1997 Dec;19(6):1225-35 PMID: 9427246
  88. CXC chemokines in angiogenesis of cancer.
    Semin Cancer Biol. 2004 Jun;14(3):195-200 PMID: 15246055
  89. Fabrication of patterned multicomponent protein gradients and gradient arrays using microfluidic depletion.
    Anal Chem. 2003 Nov 1;75(21):5775-82 PMID: 14588017
  90. Interleukin 8 (monocyte-derived neutrophil chemotactic factor) dynamically regulates its own receptor expression on human neutrophils.
    J Biol Chem. 1990 Jan 5;265(1):183-9 PMID: 2403554
  91. Cell migration and metastasis as targets of small RNA-based molecular genetic analyses.
    J Muscle Res Cell Motil. 2004;25(4-5):303-8 PMID: 15548858
  92. Squeezing axons out of the gray matter: a role for slit and semaphorin proteins from midline and ventral spinal cord.
    Cell. 2000 Aug 4;102(3):363-75 PMID: 10975526
  93. A paracrine loop between tumor cells and macrophages is required for tumor cell migration in mammary tumors.
    Cancer Res. 2004 Oct 1;64(19):7022-9 PMID: 15466195
  94. Generation of stable concentration gradients in 2D and 3D environments using a microfluidic ladder chamber.
    Biomed Microdevices. 2007 Oct;9(5):627-35 PMID: 17530414
  95. Turning of retinal growth cones in a netrin-1 gradient mediated by the netrin receptor DCC.
    Neuron. 1997 Dec;19(6):1211-24 PMID: 9427245
  96. Interleukin-8 stimulates cell proliferation in non-small cell lung cancer through epidermal growth factor receptor transactivation.
    Lung Cancer. 2007 Apr;56(1):25-33 PMID: 17175059
  97. A microfluidic multi-injector for gradient generation.
    Lab Chip. 2006 Jun;6(6):764-8 PMID: 16738728
  98. Tumor necrosis factor-alpha decreases neutrophil chemotaxis to N-formyl-1-methionyl-1-leucyl-1-phenylalanine: analysis of single cell movement.
    J Leukoc Biol. 1992 Dec;52(6):630-6 PMID: 1464735
  99. Cross-desensitization of receptors for peptide chemoattractants. Characterization of a new form of leukocyte regulation.
    J Immunol. 1994 Oct 1;153(7):3267-75 PMID: 8089498
  100. Anterior-posterior guidance of commissural axons by Wnt-frizzled signaling.
    Science. 2003 Dec 12;302(5652):1984-8 PMID: 14671310
  101. Axon guidance: stretching gradients to the limit.
    Neural Comput. 1998 Apr 1;10(3):521-7 PMID: 9527831
  102. Interaction between enteroviruses and human endothelial cells in vitro. Alterations in the physical properties of endothelial cell plasma membrane and adhesion of human granulocytes.
    Am J Pathol. 1985 Jan;118(1):15-25 PMID: 2981472
  103. Microfluidic large-scale integration.
    Science. 2002 Oct 18;298(5593):580-4 PMID: 12351675
  104. The stromal derived factor-1/CXCL12-CXC chemokine receptor 4 biological axis in non-small cell lung cancer metastases.
    Am J Respir Crit Care Med. 2003 Jun 15;167(12):1676-86 PMID: 12626353
  105. Protein and cell micropatterning and its integration with micro/nanoparticles assembly.
    Biosens Bioelectron. 2007 Jan 15;22(6):775-88 PMID: 16621507
  106. Chemoattractant receptor cross-desensitization.
    J Biol Chem. 1999 Mar 5;274(10):6027-30 PMID: 10037679
  107. Polarization and directed migration of murine neutrophils is dependent on cell surface expression of CD44.
    Cell Immunol. 2004 Sep-Oct;231(1-2):146-57 PMID: 15919379
  108. An intracellular signaling hierarchy determines direction of migration in opposing chemotactic gradients.
    J Cell Biol. 2002 Oct 14;159(1):91-102 PMID: 12370241
  109. Mitogenic effects of interleukin-8/CXCL8 on cancer cells.
    Future Oncol. 2005 Oct;1(5):699-704 PMID: 16556047
  110. Morpheus unbound: reimagining the morphogen gradient.
    Cell. 2007 Jan 26;128(2):245-56 PMID: 17254964
  111. Semaphorin-neuropilin interactions underlying sympathetic axon responses to class III semaphorins.
    Neuron. 1998 Dec;21(6):1283-90 PMID: 9883722
Article Info
Journal
Lab on a chip
Abbr.
Lab Chip
ISSN
1473-0197
DOI
10.1039/b711887b
Published
2008-01-00
Epub
2007-00-06
Pages
34-57
Language
English
Region
England
NLM ID
101128948
PMCID
PMC3848882
Subset
IM
Grants
NINDS NIH HHS · R01 NS064387 · United States
NIBIB NIH HHS · R33 EB003307 · United States
External Links
PubMed source DOI: 10.1039/b711887b PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2