The main difference between crossover cables and straight-through cables lies in their wire arrangement. Crossover cables have crisscrossed pairs of wires that reverse the incoming and outgoing signals, while straight-through cables have matching wired pins. Crossover cables are commonly used for connecting devices with the same functionality, such as computers or routers, while straight-through cables are used for connecting different devices. Additionally, crossover cables follow different wiring standards, such as T568A and T568B, on each end.