Our lab is currently interested in studying the immunological effects of Taenia crassiceps larvae in an in vitro rodent hippocampal brain slice culture model. After homogenising and centrifuging the larvae in PBS you end up with three fractions: an insoluble pellet, a soluble supernatant, and an insoluble fatty layer on top.

I am searching for the best way in which to optimally solubilise the two insoluble fractions (as tegumental glycoconjugates have been implicated in the immune modulatory abilities of Taenia larvae), but need to do this using a method that preserves brain cell viability in culture.

One paper that used Taenia solium larval antigens in a cell culture setting state that they created larval antigen suspensions in PBS by sonicating the insoluble fractions at 70Hz for 3min. I can’t seem to find any confirmation of this, and am not sure whether this is a viable solution and if it is, how it allows for the solubilisation of hydrophobic elements?

Beyond this I have thought of performing the homogenisation in a minimal volume of DMSO, and then heavily diluting the resultant supernatant with PBS/cell culture media until the concentration of DMSO >/= 0.25% (and thus minimally neurotoxic), although the resultant antigen solution may then be too dilute to detect any immunological modulations.

Has anyone successfully used sonication for this type of application?

Can anyone suggest any better ways to achieve solubility?

Thanks in advance!