Induction and detection of human mesenchymal stem cell migration in the 48-well reusable transwell assay.
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.
Rüster
Brigitte
B
Institute of Transfusion Medicine and Immune Hematology, German Red Cross Blood Donor Service, University of Frankfurt, D 60528 Germany.
Grace
Bithiah
B
Seitz
Oliver
O
Seifried
Erhard
E
Henschler
Reinhard
R
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
D005353:Fibronectins; D019096:Vitronectin