[An experimental study of coculture of esophageal mucosa epithelial cells with SIS and their biological characteristics].
OBJECTIVE
To explore an effective method to cultivate esophageal mucosa epithelial cells (EMECs) of canine in vitro, and to observe the biological characteristics of EMECs growing on SIS in order to provide an experimental basis for esophagus tissue engineering.


METHODS
Esophageal tissues were obtained from five healthy dogs aged 2 to 5 weeks under sterile conditions. The primary EMECs were cultivated with defined keratinocyte serum free medium (DKSFM) containing 6% FBS. The morphological characteristics and the growth curve of EMECs of the 2nd generation were observed for 1 to 5 days. The expressions of the EMECs marker (cytokeratin 19, CK-19) were examined by immunocytochemistry. The 2nd generation of EMECs was seeded on SIS and observed by HE staining, immunohistochemical staining, and SEM for 4 and 8 days.


RESULTS
The primary culture of canine EMECs arranged like slabstone. Immunohistochemical staining of CK-19 of the 2nd generation EMECs showed positive broadly. The cells growth reached the peak level at 2 days by MTT method. EMECs were polygon in shape and arranged like slabstone, and formed a single layer on the surface of SIS. The cells were contacted closely with each other for 4 days. Eight days later, 2 to 3 layers stratified structure was formed. Lots of EMECs were grown on SIS, and showed laminate arrangement.


CONCLUSIONS
With mixed enzymatic digestion, the culture of EMECs in DKSFM containing 6% FBS is a simple and feasible method. SIS shows good biocompatibility and can be used as a good scaffold material in the tissue engineered esophagus.
Tan
Bo
B
Division of Stem Cell and Tissue Engineering, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China.
Wei
Renqian
R
Yang
Zhiming
Z
Li
Xiuqun
X
Han
Ping
P
Zhi
Wei
W
Xie
Huiqi
H
D000818:Animals; D001672:Biocompatible Materials; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D018920:Coculture Techniques; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology*; D004947:Esophagus / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*
D001672:Biocompatible Materials