Direct cell surface modification with DNA for the capture of primary cells and the investigation of myotube formation on defined patterns

Previously, we reported a method for the attachment of living cells to surfaces through the hybridization of synthetic DNA strands attached to their plasma membrane. The oligonucleotides were introduced using metabolic carbohydrate engineering, which allowed reactive tailoring of the cell surface gl...

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Veröffentlicht in:Langmuir : the ACS journal of surfaces and colloids. - 1992. - 25(2009), 12 vom: 16. Juni, Seite 6985-91
1. Verfasser: Hsiao, Sonny C (VerfasserIn)
Weitere Verfasser: Shum, Betty J, Onoe, Hiroaki, Douglas, Erik S, Gartner, Zev J, Mathies, Richard A, Bertozzi, Carolyn R, Francis, Matthew B
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2009
Zugriff auf das übergeordnete Werk:Langmuir : the ACS journal of surfaces and colloids
Schlagworte:Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. DNA Primers DNA 9007-49-2
Beschreibung
Zusammenfassung:Previously, we reported a method for the attachment of living cells to surfaces through the hybridization of synthetic DNA strands attached to their plasma membrane. The oligonucleotides were introduced using metabolic carbohydrate engineering, which allowed reactive tailoring of the cell surface glycans for chemoselective bioconjugation. While this method is highly effective for cultured mammalian cells, we report here a significant improvement of this technique that allows the direct modification of cell surfaces with NHS-DNA conjugates. This method is rapid and efficient, allowing virtually any mammalian cell to be patterned on surfaces bearing complementary DNA in under 1 h. We demonstrate this technique using several types of cells that are generally incompatible with integrin-targeting approaches, including red blood cells and primary T-cells. Cardiac myoblasts were also captured. The immobilization procedure itself was found not to activate primary T-cells, in contrast to previously reported antibody- and lectin-based methods. Myoblast cells were patterned with high efficiency and remained undifferentiated after surface attachment. Upon changing to differentiation media, myotubes formed in the center of the patterned areas with an excellent degree of edge alignment. The availability of this new protocol greatly expands the applicability of the DNA-based attachment strategy for the generation of artificial tissues and the incorporation of living cells into device settings
Beschreibung:Date Completed 17.08.2009
Date Revised 11.05.2024
published: Print
Citation Status MEDLINE
ISSN:1520-5827
DOI:10.1021/la900150n