Microinjection for the ex Vivo Modification of Cells with Artificial Organelles

Microinjection is extensively used across fields to deliver material intracellularly. Here we address the fundamental aspects of introducing exogenous organelles into cells to endow them with artificial functions. Nanocarriers encapsulating biologically active cargo or extreme intraluminal pH were injected directly into the cytosol of cells, where they bypassed subcellular processing pathways and remained intact for several days. Nanocarriers' size was found to dictate their intracellular distribution pattern upon injection, with larger vesicles adopting polarized agglomerated distributions and smaller colloids spreading evenly in the cytosol. This in turn determined the symmetry or asymmetry of their dilution following cell division, ultimately affecting the intracellular dose at a cell population level. As an example of microinjection's applicability, a cell type relevant for cell-based therapies (dendritic cells) was injected with vesicles, and its migratory properties were studied in a co-culture system mimicking lymphatic capillaries.