BACKGROUND:
Wharton's Jelly-derived
mesenchymal stem cells are relatively primitive
stem cells that are ideal vectors for
gene therapy. However, there is a lack of studies on the conditions for the electrotransfection of
Wharton's Jelly-derived
mesenchymal stem cells. Therefore, exploring the optimal conditions for the electrotransfection of
Wharton's Jelly-derived
mesenchymal stem cells occupies an important position.
OBJECTIVE:
To investigate the effects of different
electroporation conditions on the
transfection efficiency of
Wharton's Jelly-derived
mesenchymal stem cells, and to explore the optimal conditions for
cell electroporation.
METHODS:
By controlling the
transfection conditions such as voltage,
pulse duration and
cell status, EEV-EGFP
plasmids were transfected into
Wharton's Jelly-derived
mesenchymal stem cells by
electroporation under different conditions.
Transfection efficiency was detected by
flow cytometry. RESULTS AND
CONCLUSION:
(1) The
transfection efficiency was intended to increase when the voltage ranged from 125 V to 150 V, and the maximum
transfection efficiency was obtained when the voltage was 150 V. However, when the voltage was further increased to 170 V, the
transfection efficiency began to decrease considerably. (2) The maximum
transfection efficiency was obtained when the
pulse duration was 5.0 ms, while it was certainly decreased when the
pulse duration was 2.5 and 7.5 ms. (3) The
transfection efficiency of the
cells cultured under normoxia was higher than that under hypoxic
culture. These findings reveal that normally cultured
Wharton's Jelly-derived
mesenchymal stem cells can achieve higher
electroporation efficiency via two
pulse sessions at a voltage of 150 V,
pulse duration of 5.0 ms, and
pulse interval
time of 50 ms.