Tick cell lines have emerged as a valuable tool in many research areas concerning ticks and tick-borne diseases. This has established a useful model for investigating interactions between ticks and pathogenic agents, as various economically significant tick-borne pathogens can be isolated and propagated in tick cell lines. However, tick cell lines are relatively fragile compared to more common cell types and demand higher maintenance culture conditions. Here, we describe the basic methods for tick cell culture using ISE6, a cell line derived from the embryos of the Ixodes scapularis tick species, to showcase its morphological features. Moreover, this approach can be adapted for in vitro cultivation of viral and bacterial pathogens.