title: Lentiviral transduction of rat Sertoli cells as a means to modify gene expression.
abstract: Primary cell culture is an established and widely used technique to study Sertoli cell function in vitro. However, the relative difficulty of stably overexpressing or knocking down genes in Sertoli cell culture has limited progress in the field. In this technical report, we present a method to transduce 20 dpp rat Sertoli cell cultures with VSV-G pseudotyped lentiviral based vectors at a high rate (~80%), with stable reporter gene expression. Although high transgene expression is desirable, it was noted that at transduction rates > 60% inter-Sertoli cell tight junction integrity and, hence, Sertoli cell function, were transiently compromised. We envisage that this optimized procedure has the potential to stimulate Sertoli cell research, and motivate the use of Sertoli cells in various cell therapy applications.
authors.affiliation: Prince Henry's Institute of Medical Research; Clayton, VIC Australia ; Department of Biochemistry and Molecular Biology; Monash University; Clayton, VIC Australia.
authors.forename: Peter K
authors.initials: PK
authors.lastname: Nicholls
authors.forename: Peter G
authors.initials: PG
authors.lastname: Stanton
authors.forename: Katarzyna E
authors.initials: KE
authors.lastname: Rainczuk
authors.forename: Hongwei
authors.initials: H
authors.lastname: Qian
authors.forename: Paul
authors.initials: P
authors.lastname: Gregorevic
authors.forename: Craig A
authors.initials: CA
authors.lastname: Harrison