Inspect the pumpkin vine to make sure it is producing female flowers, or those with immature or rudimentary fruit beneath the flower petals. Pumpkin vines will produce many male flowers before the female flowers begin to develop to make sure pollen is readily available for fertilization.
Break off a mature male flower -- one with abundant pollen that comes off on your hand when you touch its stamen or pollen-bearing structure. Cut the male flower's petals off so the stamen is open to the air.
Find a freshly opened female flower. Female flowers are receptive to fertilization for only one day, so it's important that the flower is fresh or newly opened.
Roll the male flower's pollen onto the female flower's stigma, in the center part of the flower, making sure that pollen covers all stigma sections.