Biomedical Engineering Reference
In-Depth Information
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Figure 11.6
Cartoon of cell-accumulation technique by FN-G nanofilm
fabrication (A). HE staining image of 23L-NHDF tissues after 24 h of
incubation (B).
Reproduced with permission from Ref. 85.
fluorescently labeled FN-G nanofilms onto the surface of dispersed cells.
Furthermore, we also confirmed over 91% cell viability for NHDFs,
HUVECs, and HepG2 cells after preparing the FN-G films. After coating with
the FN-G nanofilms, 2
10
6
NHDFs were incubated in a cell culture insert
for one day. The 8L-dense constructs of 35
4 mm thicknesses were suc-
cessfully obtained, whereas a tattered and porous structure was barely ob-
tained using noncoated cells. The thickness of the obtained tissue
constructs increased on increasing the volume of culture media, and thus
the current maximum thickness was over 100 mm, more than 20L structures
(Figure 11.6B). More importantly, the construction of approximately 20L
structures per one day after only a nanofilm coating within 1 h has never
been reported previously, and this approach has tremendous versatility for
various cells.
We demonstrated fabrication of vascularized 3D-tissue arrays were con-
structed in 12 and 24 microwell plates using a cell-accumulation techni-
que.
85
The construction of the thick multilayered tissues with endothelial
tube networks by embedding HUVECs in 3D-tissues composed of NHDFs
has been performed by sandwich culture. After 2-7 days of incubation,
highly developed capillary networks and a tubular morphology of the
HUVECs were clearly observed by CLSM analyses (Figure 11.7). A dense and
homogeneous vascularized network in the multilayered tissues of 1 cm
width and 50 mm height was confirmed in 12 and 24 microwell cell-culture
inserts. The occupied area percentage and distance of this capillary network
of HUVECs was calculated to be approximately 63
12% and 50-150 mmby
.
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