CHITIN AND CHITOSAN FOR TISSUE ENGINEERING APPLICATION - Intelligent Scaffolds for Tissue Engineering and Regenerative Medicine - page 449

concentration and decreased with increasing of chitosan concen-
tration. The inserted image in Fig. 23.8 shows the chitosan/ β -GP
hydrogel at 37 ◦ C.

23.5.2 Cell Culture on Chitosan Hydrogels
Primary HDFs from newborn foreskin (HDF-N, MCTT Co., Korea) at
passage 6 were used. The chitosan was dissolved in 0.1M HCl to
give a 2.0% (w/v) solution, and the β -GP was dissolved in water
to give a 45% (w/v) solution. The chitosan, β -GP solution, and the
media containing HDFs were mixed by a volume ratio of 1:0.8:0.2,
respectively. The seeding cell density was 5 × 10 5 cells per 100 μ L
of thermo-gelling solution. The inoculated HDFs were cultured in
a medium (DMEM:F12 = 3:1) including 10% FBS in an incubator
under 95% humidity at 37 ◦ C with 5% CO 2 . Figure 23.9 shows
that the left and right images are a thermo-gelling hydrogel and
hematoxylin and eosin (H&E) staining of a hydrogel cultured on
HDFs for three days of culture, respectively. The result of H&E
staining showed that the cultured HDFs well distributed in the
hydrogel.
The optimum cell concentration in the chitosan/

β

-GP hydrogel

×

10 5

∼ ×

10 6 cells per 100

μ

was measured by 5 1 Lofthermo-

gelling solution.

Figure 23.9. The thermo-gelling hydrogel cultured on primary HDFs (left
image)andH&EstainingofthehydrogelculturedonHDFsfor3dofculture
(right image).