The women said the crossing needed to be reopened so that thousands of Palestinian medical patients could seek medical treatment outside the Gaza Strip, which has been suffering an all-out Israeli blockade since 2007.
The protesters carried placards on which they called for reopening the crossing and complained against the inability of Gaza's patients to travel to seek medical treatment abroad.
"Palestinians have the right to freely move through the crossing," Esraa al-Areer, the spokeswoman of "Women against the Blockade", the Palestinian movement that organized the protest, told The Anadolu Agency.
She called on Egyptian authorities to reopen the crossing to ease the travel of thousands of Gaza's Palestinians.
Al-Areer said around 38,000 Palestinians needed to travel from Gaza to other parts outside of it.
She called on Arab and Islamic states to support the Palestinians of the Gaza Strip and help them to break the Israeli siege imposed on the Palestinian territory since 2007.
Egypt closed the Rafah border crossing, Gaza's only access to the outside world that is not controlled by Israel, on October 24 following an attack on a military site in the Sinai Peninsula that left 31 Egyptian troops dead.
Egypt has tightened its grip on the border with the blockaded Gaza Strip since last year's ouster of Mohamed Morsi – the country's first freely elected president – by the army.
Repeated crossing closures have made life even more difficult for Gaza's roughly 1.9 million residents.