Establishment of primary transgenic human airway epithelial cell cultures to study respiratory virus – host interactions

Hulda R. Jonsdottir; Sabrina Marti; Dirk Geerts; Regulo Rodriguez; Volker Thiel; Ronald Dijkman

In an initial effort to establish transgenic hAEC cultures, we attempted to transduce 234 well-differentiated cultures directly with vesicular stomatitis virus glycoprotein (VSV-g) pseudotyped 235 lentiviral particles harboring a pLKO_GFP transfer vector. However, intact cultures are refractory to 236 both apical and basolateral transduction while damaged cultures can be transduced from the apical 237 side when the basolateral side of the epithelium is exposed. In those cultures, GFP expression is only 238 observed around the edges of damaged epithelium (data not shown). This is consistent with the 239 observation that receptors for VSV-g are predominantly located on the basolateral side of polarized The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/694380 doi: bioRxiv preprint Figure 2B) . Thus, the removal of the inhibitor from culture medium upon LLI 307 culture seems to be sufficient for the cells to proceed through differentiation normally post-treatment.