Site directed adhesion of cells can be obtained by cultivation of adherent cells on patterned substrates containing cell-repellent regions and regions where cells adhere well. Of particular interest are substrates that can be dynamically altered as a cell cultivation experiment proceeds [1-3], for instance in order to introduce new cell types into previously cultured cells.

Patterned oligoethyleneglycol-terminated self-assembled monolayers (OEG SAMs) were created by microelectrochemically formation of Br₂ [4-6]. These substrates were used to monitor local adherence of astrocyts derived from the brains of newborn rats [7] and oligodendroglia OLN-93 cells [8]. These cells adhere without other specific substrate proteins and occupy selectively the manipulated OEG SAM surface.

Bromide is necessary as precursor for the microelectrochemical Br₂ formation used for the modification of the OEG SAM. For the sequential introduction of cells into pre-existing patterns, it is important that adherent cells can withstand bromide concentrations in the culture medium. For that reason, the cytotoxicity of KBr addition to the culture medium (Dulbecco's Modified Eagle's Medium with 10 % fetal calf serum) was tested for OLN-93 cells between 25-100 mM KBr. The results show that these cells can tolerate 50 mM KBr concentration for the duration of 2 hours.

For sequential co-culture experiments on patterned substrates, two clonal cell lines OLN-93 are used: OLN-93 cells transfected with GFP protein and OLN-93 cells transfected with DsRed protein. The local attachment of the different cell populations could be detected by fluorescence microscopy like confocal laser scanning microscopy (CLSM).

References

1.   M. N. Yousaf, B. T. Houseman, M. Mrksich, Proc. Natl. Acad. Sci. U.S.A. 2001, 98, 5992-5996.      

2.   X. Jiang, R. Ferrigno, M. Mrksich, G. M. Whitesides, J. Am. Chem. Soc. 2003, 125, 2366-2367.

3.   Y. Li, B. Yuan, H. Ji, D. Han, S. Chen, F. Tian, X. Jiang, Angew. Chem. Int. Ed. 2007, 46, 1094-1096.

4.   C. Zhao, I. Zawisza, M. Nullmeier, M. Burchardt, M. Träuble, I. Witte, G. Wittstock, Langmuir 2008, 24, 7605-7613.

5.   C. Zhao, I. Witte, G. Wittstock, Angew. Chem. Int. Ed. 2006, 45, 5469-5471.

6.   I. Brand, M. Nullmeier, R. Jogireddy, J. Christoffers, G. Wittstock, Langmuir 2009, in press.

7.   O. Goldbaum, G. Vollmer, C. Richter-Landsberg, Glia 2006, 53, 891-901.

8.   C. Richter-Landsberg, M. Heinrich, J. Neuroscience Res. 1996, 45, 161-173.

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