In Vitro Testing of Anticancer Nanotherapeutics Using Tumor Spheroids.

Tumor cells grown as spheroids more closely resemble in vivo solid tumors and present physical and physiological barriers to drug action that conventional monolayer cell cultures do not. The physiological relevance of the model can be further increased by incorporating fluid flow and multiple cell types. The protocols described in this chapter use simple and easily available equipment to reproducibly generate spheroid models that can be used for the in vitro testing of a variety of preparations.

Synthesis of triphenylphosphonium phospholipid conjugates for the preparation of mitochondriotropic liposomes.

Surface modification of liposomes with a ligand is facilitated by the conjugation of the ligand to a hydrophobic molecule that serves to anchor the ligand to the liposomal bilayer. We describe here a simple protocol to conjugate a triphenylphosphonium group to several commercially available functionalized phospholipids. The resulting triphenylphosphonium conjugated lipids can be used to prepare liposomes that preferentially associate with mitochondria when exposed to live mammalian cells in culture.