[Culture and purification of human fetal OB olfactory ensheathing cells in vitro with different methods].
OBJECTIVE
To explore simple and useful methods for primary culture of olfactory ensheathing cells (OECs), which could harvest human fetal OB olfactory ensheathing cells (OB-hOECs) with high purity in vitro.


METHODS
OB-hOECs were cultured and purified with different methods. The basic method was dependent on the differing rates of cell attachment, the other methods were combined the basic method with cytarabine method, starvation method without serum, and NT3, method without serum respectively. The growth of OECs was observed and the purity of cultured cells was identified with immunocytochemistry.


RESULTS
OB-hOECs appears dipolar and tripolar shape, and have slender mess evection forming network. The purity of OECs reached 88% with the basic method in 6 div, then reduced quickly due to fibroblast proliferation. With the starvation method, the OECs reached a higher purity of 91% at 6 div and 86% at 9 div, but cell condition was not very well. For the method of NT3 without serum, the purity could touch the highest score 95% at 9 div, and 83% at 12 div with a good cell condition. The purity of OECs cultured by the method of NT, without serum is statistical significantly higher than that of other methods (P<0.05).


CONCLUSIONS
Associated with the basic method of different cell attachment rate, the NT3 without serum method could obtain OECs with higher purity and better cell condition.
Wang
Bin
B
Department of Orthopedics, Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710004, China.
He
Xi-jing
XJ
Li
Qiang
Q
Li
Hao-peng
HP
Wang
Dong
D
D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D003561:Cytarabine / Q000494:pharmacology; D005333:Fetus; D006801:Humans; D020933:Neurotrophin 3 / Q000494:pharmacology; D009830:Olfactory Bulb / Q000166:cytology* / Q000196:embryology
D016895:Culture Media, Serum-Free; D020933:Neurotrophin 3; D003561:Cytarabine