[Reconstruction of rabbit corneal stroma with skin fibroblasts].
OBJECTIVE
To explore whether skin fibroblasts could be used as a cell source for reconstruction of the corneal stroma.


METHODS
It was an experimental study. Skin fibroblast cells were isolated from newborn rabbits, cultured and expanded in vitro. Cells were labeled with green fluorescence protein (GFP) gene by retro-viral infection. Fibroblasts at passage 3 were seeded on polyglycolic acid (PGA) non-woven fibers to form a cell-scaffold construct. Constructs were then implanted into the adult rabbit corneal stroma layer after being cultured in vitro for 1 week. Engineered stroma were observed continuously and harvested after 8 weeks of transplantation for gross, histological evaluation and Keratocan examination. PGA alone was used as control.


RESULTS
The engineered tissue in the cornea became transparent gradually over a period of 8 weeks. Histological analysis showed that engineered stromal lamellar was relatively regular and the orientation of fibers was parallel to the surface of cornea, which is similar to normal cornea. The implanted cells were confirmed by GFP expression under fluorescent microscope, which also express Keratocan. By transmission electron microscopy examination, no significant difference in the diameter of collagen fiber was observed between engineered stroma (33.08 + or - 2.47) nm and normal stroma (t = 1.80, P = 0.0771).


CONCLUSIONS
Skin fibroblast cells could be used as seed cells for reconstruction of the corneal stroma.
Department of Ophthalmology, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai 200031, China.
Yan-qing
YQ
Zhang
Wen-jie
WJ
Zhang
Wei
W
Liu
Xiao-jie
XJ
Hu
Guang-dong
GD
Zhou
Lei
L
Cui
Yi-lin
YL
Cao
D000818:Animals; D002478:Cells, Cultured; D003319:Corneal Stroma; D005109:Extracellular Matrix; D005347:Fibroblasts / Q000166:cytology*; D011817:Rabbits; D012867:Skin / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds