FK506 promoting proliferation of Schwann cells in vitro and NGF of Schwann cells secreted highly by itself
- VernacularTitle:FK506对许旺细胞体外增殖和分泌NGF的影响
- Author:
Chunhua LI
;
Jun YANG
;
Lei WU
;
Jianwen LIAO
;
Zehua CHEN
;
Qiping WANG
;
Zhenwei ZHANG
;
Jianqiang QIN
- Publication Type:Journal Article
- Keywords:
FK506;
Sehwann cell;
NGF;
Peripheral nerve;
Tissue engineering
- From:
Chinese Journal of Microsurgery
2008;31(6):428-431,illust 4
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore on FK506 promoting proliferation of Schwann cells in vitro and NGF of Schwaun cells secreted highly by itself. Methods Purified Schwann cells divide into six groups:group A (control group) DMEM/F12 contained 10% calf bloodserum; group B contained 0.1 ng/ml FK506; group C contained 0.5 ng/ml FK506; group D contained FK506:1.0 ng/ml;group E contained FK506:5.0 ng/ml; group F contained FK506:10 ng/ml. Morphology of Schwann cells were oboyrved by invert microscope and evaluated Schwann cells in immunocytochemistry staining with anti-S-100. The best concentration of FK506 who promoted proliferation of Schwann cells by MTT. Cell cycle of Sehwarm cells were determined by flow cytometry. The level of NGF in the conditioned media was determined by an enzyme-linked immunoadsordcnt assay after 72 h. Results Group C was the best concentration which promoting proliferation of Schwann cells among 5 groups, when the concentration 1.0 ng/ml FKS06 to promote Schwann cell proliferation gradually weakened. Detected by flow cytometry showed that: containing 10% fetal DMEM/F12 bovine serum for 24 h,72 h and 48 h after Schwann cells in S phase percentage were 27.8%,39.3% and 58.4% in the 0.5 ng / ml FK506 for 24 h,72 h and 48 h after Schwann cells S percentage period were 54.2% ,60.3% and 94.6%. S phase of the latter than the former in 24 h,72 h and 48 h, respectively higher: 26.4% and 21% and 36.2%. FK506 detected by ELISA promote Schwann cell proliferation after the expression of NGF in the experimental study found: 0.5 ng/ml FK506 for 72 h after the Schwann cells secreted by the NGF as high as 0.188 ng/ml, rcspectiveIy. Conclusion FKS06 can promote proliferation of Schwann cells at early time in vitro and Schwann cells' good situation is highly kept to secrete NGF.