Cell adhesion profiling using extracellular matrix protein microarrays.
We have developed a microarray-based system for cell adhesion profiling of large panels of cell-adhesive proteins to increase the throughput of in vitro cell adhesion assays, which are currently primarily performed in multiwell plates. Miniaturizing cell adhesion assays to an array format required the development of protocols for the reproducible microspotting of extracellular matrix (ECM) protein solutions and for the handling of cell suspensions during the assay. We generated ECM protein microarrays with high reproducibility in microspot protein content using nitrocellulose-coated glass microslides, combined with piezoelectric microspotting of protein solutions. Protocols were developed that allowed us to use 5000 cells or fewer on an array of 4 x 4 mm consisting of 64 microspots. Using this microarray system, we identified differences of adhesive properties of three cell lines to 14 different ECM proteins. Furthermore, the sensitivity and accuracy of the assays were increased using microarrays with ranges of ECM protein amounts. This microarray system will be particularly useful for extensive comparative cell adhesion profiling studies when only low amounts of adhesive substrate and cells, such as stem cells or cells from biopsies, are available.
Kuschel
Cornelia
C
NMI at the University of Tübingen, Reutlingen, Germany.
Steuer
Heiko
H
Maurer
Andreas N
AN
Kanzok
Britta
B
Stoop
Reinout
R
Angres
Brigitte
B
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000032:analysis*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D016326:Extracellular Matrix Proteins / Q000032:analysis*; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007668:Kidney / Q000378:metabolism* / Q000502:physiology*; D051379:Mice; D041681:NIH 3T3 Cells; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*
D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins