[Effects of epithelial cell conditioned medium on differentiation of BMSCs].
OBJECTIVE
To investigate the feasibility of inducing canine BMSCs to differentiate into epithelial cells in vitro with epithelial cell conditioned medium (ECCM).


METHODS
Five mL BMSCs were obtained from iliac spine of a healthy adult male canine with weighing 10 kg, and then isolated and cultured. The oral mucosa was harvested and cut into 4 mm x 4 mm after the submucosa tissue was eliminated; ECCM was prepared. BMSCs of the 2nd passage were cultured and divided into two groups, cultured in ECCM as experimental group and in L-DMEM as control group. The cell morphological characteristics were observed and the cell growth curves of two groups were drawn by the continual cell counting. The cells were identified by immunohistochemical staining through detecting cytokeratin 19 (CK-19) and anti-cytokeratin AE1/AE3 on the 21st day of induction. The ultra-structure characteristics were observed under transmission electron microscope.


RESULTS
The cells of two groups showed long-fusiform in shape and distributed uniformly under inverted phase contrast microscope. The cell growth curves of two groups presented S type. The cell growth curve of the experimental group was right shifted, showing cell proliferation inhibition in ECCM. The result of immunohistochemical staining for CK-19 and anti-cytokeratin AE1/AE3 was positive in the experimental group, confirming the epithelial phenotype of the cells; while the result was negative in the control group. The cells were characterized by tight junction under transmission electron microscope.


CONCLUSIONS
The canine ECCM can induce allogenic BMSCs to differentiate into epithelial cells in vitro.
Yang
Peng
P
Division of Stem Cell and Tissue Engineering, National Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R. China.
Wei
Renqian
R
Tan
Bo
B
Li
Xiuqun
X
Wang
Jia
J
Zhou
Kunpeng
K
Zuo
Xiao
X
Li
Shun
S
Xie
Huiqi
H
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology*; D008297:Male; D013234:Stem Cells / Q000166:cytology*; D023822:Tissue Engineering
D017077:Culture Media, Conditioned