title: Induction and detection of human mesenchymal stem cell migration in the 48-well reusable transwell assay.
abstract: The induction and detection of chemotactic migration in mesenchymal stem cells (MSC) are complicated by their adherent nature. We describe here the experimental details for inducing the transmigration of MSC seeded in microchemotaxis chambers and reliably identifying the translocated cells. A combination of 8-microm pore-sized membranes and a two-step staining procedure resulted in the detection of MSC migration against a gradient of human plasma which plateaued after 4 h. Microscopic enumeration of the transmigrated cells clearly distinguished contaminating nonmigrated cells from transmigrated cells. Chemokinetic and chemotactic effects could be separated and were influenced by precoated fibronectin or vitronectin. Thus, the technique described allows rapid and reliable induction and determination of migration in adherent MSC.
authors.affiliation: Institute of Transfusion Medicine and Immune Hematology, German Red Cross Blood Donor Service, University of Frankfurt, D 60528 Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Rüster
authors.forename: Bithiah
authors.initials: B
authors.lastname: Grace
authors.forename: Oliver
authors.initials: O
authors.lastname: Seitz
authors.forename: Erhard
authors.initials: E
authors.lastname: Seifried
authors.forename: Reinhard
authors.initials: R
authors.lastname: Henschler
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002465:Cell Movement; D002633:Chemotaxis; D004305:Dose-Response Relationship, Drug; D005353:Fibronectins / Q000737:chemistry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods; D019096:Vitronectin / Q000737:chemistry
chemical_list: D005353:Fibronectins; D019096:Vitronectin