Lentiviral transduction of rat Sertoli cells as a means to modify gene expression.
Peter K Nicholls, Peter G Stanton, Katarzyna E Rainczuk, Hongwei Qian, Paul Gregorevic, Craig A Harrison
Spermatogenesis | Published : 2012
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..View full abstract