title: A novel three-dimensional flow chamber device to study chemokine-directed extravasation of cells circulating under physiological flow conditions.
abstract: Extravasation of circulating cells from the bloodstream plays a central role in many physiological and pathophysiological processes, including stem cell homing and tumor metastasis. The three-dimensional flow chamber device (hereafter the 3D device) is a novel in vitro technology that recreates physiological shear stress and allows each step of the cell extravasation cascade to be quantified. The 3D device consists of an upper compartment in which the cells of interest circulate under shear stress, and a lower compartment of static wells that contain the chemoattractants of interest. The two compartments are separated by porous inserts coated with a monolayer of endothelial cells (EC). An optional second insert with microenvironmental cells of interest can be placed immediately beneath the EC layer. A gas exchange unit allows the optimal CO2 tension to be maintained and provides an access point to add or withdraw cells or compounds during the experiment. The test cells circulate in the upper compartment at the desired shear stress (flow rate) controlled by a peristaltic pump. At the end of the experiment, the circulating and migrated cells are collected for further analyses. The 3D device can be used to examine cell rolling on and adhesion to EC under shear stress, transmigration in response to chemokine gradients, resistance to shear stress, cluster formation, and cell survival. In addition, the optional second insert allows the effects of crosstalk between EC and microenvironmental cells to be examined. The translational applications of the 3D device include testing of drug candidates that target cell migration and predicting the in vivo behavior of cells after intravenous injection. Thus, the novel 3D device is a versatile and inexpensive tool to study the molecular mechanisms that mediate cellular extravasation.
authors.affiliation: Torrey Pines Institute for Molecular Studies.
authors.forename: Valentina
authors.initials: V
authors.lastname: Goncharova
authors.forename: Sophia K
authors.initials: SK
authors.lastname: Khaldoyanidi
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002465:Cell Movement / Q000502:physiology*; D018925:Chemokines / Q000494:pharmacology*; D003584:Cytological Techniques / Q000295:instrumentation* / Q000379:methods; D042783:Endothelial Cells / Q000166:cytology*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D051379:Mice
chemical_list: D018925:Chemokines
references.citation: Cell Stem Cell. 2008 Mar 6;2(3):198-200
references.pmid: 18371444
references.citation: ASGSB Bull. 1991 Jul;4(2):87-94
references.pmid: 11537186
references.citation: J Leukoc Biol. 2001 Jun;69(6):860-6
references.pmid: 11404368
references.citation: J Clin Invest. 1993 Dec;92(6):3038-44
references.pmid: 7504697
references.citation: J Biol Chem. 2003 Feb 7;278(6):4127-34
references.pmid: 12438311
references.citation: J Exp Med. 1998 Aug 3;188(3):465-74
references.pmid: 9687524
references.citation: Cell Stem Cell. 2012 Jun 14;10(6):740-749
references.pmid: 22704514
references.citation: Curr Stem Cell Res Ther. 2008 Dec;3(4):229-36
references.pmid: 19075753
references.citation: Biol Blood Marrow Transplant. 2012 Sep;18(9):1407-14
references.pmid: 22430086
references.citation: Am J Physiol Lung Cell Mol Physiol. 2001 Sep;281(3):L529-33
references.pmid: 11504676
references.citation: Ann Biomed Eng. 2012 Apr;40(4):766-76
references.pmid: 22068886
references.citation: Hematology Am Soc Hematol Educ Program. 2010;2010:1-6
references.pmid: 21239763
references.citation: Circ Res. 2008 May 23;102(10):1155-68
references.pmid: 18497316
references.citation: Mol Med Today. 1999 Jan;5(1):40-6
references.pmid: 10088131
references.citation: Int J Mol Med. 1999 Oct;4(4):323-32
references.pmid: 10493972
references.citation: Cell. 2008 Feb 22;132(4):612-30
references.pmid: 18295579
references.citation: J Immunol. 2009 Mar 15;182(6):3827-36
references.pmid: 19265162
references.citation: Bone Marrow Transplant. 2011 May;46(5):650-8
references.pmid: 20711241
references.citation: Methods Mol Biol. 2011;769:331-49
references.pmid: 21748686