Related Experiment Video
Updated: May 13, 2026
Enrichment of Pachytene Spermatocytes and Spermatids from Mouse Testes Using Standard Laboratory Equipment
Published on: September 17, 2019
[Construction and identification of a mouse spermatocyte-derived cell line with a stable expression of PIAS-NY]
Ying Zheng1, Hai-Yan Wang, Lu-Ping Zhang
1Department of Anatomy, Histology and Embryology, The Affiliated Hospital of Yangzhou University School of Medicine, Yangzhou, Jiangsu 225001, China. yzzkl@163.com
Objective:
To construct a lentiviral expression vector of the PIAS-NY gene, and establish a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.
Methods:
PIAS-NY was synthesized, amplified by PCR and cloned into the lentiviral vector expression plasmid pGC-FU. After digestion and sequencing, pGC-FU-PIAS-NY, pHelper 1.0 and pHelper 2.0 were co-transfected into 293T cells. Then the lentiviral particles were used to transfect the mouse spermatocyte-derived cells. The expression of the PIAS-NY protein was detected by Western blot.
Results:
We successfully constructed the lentiviral expression vector pGC-FU-PIAS-NY and established a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.
Conclusion:
The construction of the lentiviral expression vector pGC-FU-PIAS-NY and the obtainment of stably transfected mouse spermatocyte-derived cells have paved the way for further studies on the roles of the PIAS-NY gene in spermatogenesis.
