Home > Browse Issues > Vol.37 No.8
Characterization of Sertoli Cells from Prepubertal Porcine Testis In Vitro
Liu Huanhuan, Yu Shumin*, Xia Juan, Sun Chengliang, Shen Liuhong, Cao Suizhong, Zuo Zhicai, Ma Xiaoping
College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
Abstract: To analysis the characterization of sertoli cells in vitro， the modified digestion and differential plating methods were used for isolating sertoli cells from 3~5 week-old porcine testis. Then， cell proliferation， the expression of markers and secretion of cytokines were detected. The results showed that prepubertal porcine treated with 0.25% collagenase IV and 0.1% hyaluronidase for 60 min， and then with 0.25% trypsin and 0.1% DNase I for 20 min was the optimal method for isolating sertoli cells. And after inoculation， the adhesion process of sertoli cells were accomplished within 3~5 h. The purified cells were showed with large body and fusiform or irregular shape，and were positive for GATA4， but negative for alkaline phosphatase (CAKP) staining. Simultaneously， RT-PCR analysis showed these cells expressed GATA4， SOX9 and INHB， did not express CYP17， 3β-HSD and α-SMA. And detected by ELISA， cells express GDNF， LIF， CSF-1， EGF and FGF2. In summary， all these features were in line with sertoli cells.
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