Biology Reference
In-Depth Information
since it is entirely automated and allows continuous monitoring of the cellular
response to wounding. We have used the ECIS system to measure the influence
of the HCMV secretome on WH. For these assays, primary HUVECs were plated
in two arrays in complete medium and incubated overnight to allow establishment
of a confluent monolayer. The next day, cells were serum-starved prior to the
addition of secretome samples from mock- and AD169- (plus or minus UV or
foscarnet treatments) infected or HSV-1-infected HFs to duplicate wells. SFM
and complete medium were used as negative and positive controls, respectively.
Immediately after application of supernatants, arrays were placed in the electrode
holders and impedance was measured for a few minutes. This confirmed the
existence of confluent cell monolayers over each electrode and verified that the
cells were healthy. All but one of the chambers of cells were wounded for 30 s.
As expected, the electrical wounding resulted in an immediate drop in impedance
to the level of an open electrode. The subsequent rise in impedance due to migra-
tion and repopulation of the wound was monitored. As shown in Fig. 2 under the
presence of complete medium, the wound was repopulated in about 6 h. Similarly,
Fig. 2
HCMV secretome mediates EC wound healing. Wound healing activity of the HCMV
secretome. ECs were grown to confluence on ECIS arrays and exposed to test supernatants prior
to electrical wounding. Wound healing, as indicated by increasing resistance, is plotted as a func-
tion of time. Healing traces for duplicate wells are shown for the HSV-1 Secretome (
yellow
), mock
(
green
), HCMV (
red
), or UV-inactivated HCMV (
light blue
). Control traces include a negative
control (SSFM;
pink
), a positive control (Complete SFM;
dark blue
) and an unwounded control
(
black
). Cells exposed to the HCMV secretome show wound repair within 6-10 h, whereas cells
exposed to the HSV-1 or mock secretomes repopulate the wound inefficiently, indicating that the
production of wound healing factors is specific for HCMV infections
