title: Successive detection of insulin-like growth factor-II bound to receptors on a living cell surface using an AFM.
abstract: In this study, we have developed a method of mechanical force detection for ligands bound to receptors on a cell surface, both of which are involved in a signal transduction pathway. This pathway is an autocrine pathway, involving the production of insulin-like growth factor-II (IGF-II) and activation of the IGF-I receptor, involved in myoblast differentiation induced by MyoD in C3H10T1/2 mouse mesenchymal stem cells. Differentiation of C3H10T1/2 was induced with the DNA demethylation agent 5-azacytidine (5-aza). The etched AFM tip used in the force detection had a flat surface of which about 10 µm(2) was in contact with a cell surface. The forces required to rupture the interactions of IGF-IIs on a cell and anti mouse IGF-II polyclonal antibody immobilized on an etched AFM tip were measured within 5 days of induction of differentiation. The mean unbinding force for a single paired antibody-ligand on a cell was about 81 pN, which was measured at a force loading rate of about 440 nN/s. The percentage of unbinding forces over 100 pN increased to 32% after 2 days from the addition of 5-aza to the medium. This method could be used in non-invasive and successive evaluation of a living cell's behavior.
authors: Research Institute for Cell Engineering (RICE), National Institute of Advanced Industrial Science and Technology (AIST), Central 4, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562, Japan.
authors: Sung-Woong
authors: SW
authors: Han
authors: Shingo
authors: S
authors: Mieda
authors: Chikashi
authors: C
authors: Nakamura
authors: Takanori
authors: T
authors: Kihara
authors: Noriyuki
authors: N
authors: Nakamura
authors: Jun
authors: J
authors: Miyake
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001374:Azacitidine / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D003584:Cytological Techniques; D007335:Insulin-Like Growth Factor II / Q000378:metabolism*; D051379:Mice; D018625:Microscopy, Atomic Force; D017526:Receptor, IGF Type 1 / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D007335:Insulin-Like Growth Factor II; D017526:Receptor, IGF Type 1; D001374:Azacitidine