title: T cell activation on a single-cell level in dielectrophoresis-based microfluidic devices.
abstract: The gentle and careful in vitro processing of live cells is essential in order to make them available to future therapeutic applications. We present a protocol for the activation of single-T cells based on the contact formation with individual anti-CD3/anti-CD28 presenting microbeads in a lab-on-chip environment. The chips consist of microfluidic channels and microelectrodes for performing dielectrophoretic manipulation employing a.c. electric fields. The dielectrophoretic guiding elements allow the assembly of cell-bead pairs while avoiding ill-defined physical contacts with their environment. After overnight cultivation of the manipulated cells, 77% of the bead-associated T cells expressed the activation marker molecule CD69. Physiological stress on the cells was shown to be mainly due to the single-cell cultivation and not to the manipulation in the chips. The same approach could be useful for the in vitro regulation of stem cell differentiation.
authors.affiliation: Fraunhofer Institute for Biomedical Engineering (IBMT), Lab-On-Chip Technologies, Am Muehlenberg 13, 14476 Potsdam, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Kirschbaum
authors.forename: Magnus Sebastian
authors.initials: MS
authors.lastname: Jaeger
authors.forename: Tim
authors.initials: T
authors.lastname: Schenkel
authors.forename: Tanja
authors.initials: T
authors.lastname: Breinig
authors.forename: Andreas
authors.initials: A
authors.lastname: Meyerhans
authors.forename: Claus
authors.initials: C
authors.lastname: Duschl
mesh_terms: D000906:Antibodies / Q000494:pharmacology; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D018106:CD28 Antigens / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D002478:Cells, Cultured; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D037181:Lectins, C-Type; D008213:Lymphocyte Activation / Q000187:drug effects; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000099192:CD69 Antigens
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D018106:CD28 Antigens; D017252:CD3 Complex; D037181:Lectins, C-Type; D000099192:CD69 Antigens