4.4 Article

Efficient electroporation of peptides into adherent cells: investigation of the role of mechano-growth factor in chondrocyte culture

Journal

BIOTECHNOLOGY LETTERS
Volume 33, Issue 5, Pages 883-888

Publisher

SPRINGER
DOI: 10.1007/s10529-010-0508-1

Keywords

Chondrocytes; In situ electroporation; Indium tin oxide; Mechano-growth factor; Peptide therapeutics

Funding

  1. AO Research Foundation
  2. Swiss National Science Foundation [PMPDP2_122997]
  3. Leda Raptis
  4. Kevin Firth
  5. Tom Hole
  6. Swiss National Science Foundation (SNF) [PMPDP2_122997] Funding Source: Swiss National Science Foundation (SNF)

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Peptide therapeutics are of increasing interest due to their biological specificity. We used a simple technique to study the efficacy of inducing peptides into adherent chondrocytes by transiently permeabilizing the membrane with electric pulses (in situ electroporation). Mechano-growth factor (MGF) was selected as a model peptide. FITC-labeled MGF was added to cultures of adherent primary chondrocytes grown on ITO coated glass slides. Cells were subjected to 3-9 pulses of 175-275 V and evaluated by flow cytometry. Under optimal conditions, an electroporation efficiency of close to 50% could be achieved. This technique can be used to study the functional domains of intracellular peptides, peptide inhibition of signal transduction and intracrine-mediated effects of peptides in adherent cells.

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