Adeno-associated viral vector-mediated gene delivery and shRNA knockdown in primary human cytotrophoblasts

The placenta is essential for fetal growth and the maintenance of pregnancy. Primary human cytotrophoblasts (CTBs) provide a physiologically relevant model of this transient organ, yet their use has been limited by the difficulty of genetic manipulation. Recombinant adeno-associated virus (AAV) vectors, widely used to modulate gene expression in other systems, offer low immunogenicity, low cytotoxicity, and high efficiency in primary cells. Here, we provide proof of concept of an AAV-mediated gene delivery approach for primary CTBs. This platform enables in-depth, cell-biological studies to elucidate molecular mechanisms of human placental biology and pathology.