A crowd of well-wishers saw off the sailing vessel Estelle as it left the Italian port as part of the "Freedom Flotilla" movement.
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"We think it will take around two weeks to get to Gaza, but it will obviously also depend on the weather conditions," spokeswoman Ann Ighe said earlier.
The Estelle, whose voyage was organized by an international pro-Palestinian coalition, is carrying humanitarian goods to the Gaza Strip.
Gaza-bound vessel leaves Naples
Israel says the blockade is necessary to prevent weapons from entering the coastal territory, which is run by the Islamist movement Hamas.
The first Freedom Flotilla in May 2010 ended in tragedy when nine Turkish nationals were killed after Israeli commandos boarded the Turkish-flagged Mavi Marmara, the lead ship, as it tried to break through the blockade.
'Blockade creates bitterness and hatred.' Ship en route to Gaza (Photo: Ship to Gaza Sweden)
"When people think of flotillas, they think of the Mavi Marmara. But the Estelle is very different, she was chosen to change the perception of the campaign, to show that it's a non-violent campaign," Ighe said.
At just 53 meters long, the Estelle is much smaller than the Marmara ferry, and has sails as well as an engine.
"It is a vessel that has travelled for Greenpeace protests and on fair-trade missions. The Estelle is a peaceful ship," Ighe said.
'Estelle is a peaceful ship' (Photo: AFP)
Naples mayor Luigi de Magistris said Friday on visiting the Estelle: "This is not an initiative in favor of Hamas but in favor of Palestine, of the Palestinians who live in Gaza, and in favor of two states living as neighbors in peace and security."
Dror Feiler, who also took part in the 2010 sail to Gaza aboard the Maramara, told Ynet that the 'Estelle' is carrying 600 soccer balls, 41 tons of cement, theater equipment and musical instruments.
"We hope the Israeli government will understand that there is no point in preventing us from reaching Gaza," he said. "Our flotilla does not pose a security threat to Israel. The sail is not against the State of Israel, but against the Israeli government's policy. We believe it is completely legitimate and consider the blockade to be detrimental to both Israel and the Palestinians. It only creates bitterness and hatred."
According to Feiler, all of the activists aboard the vessel signed a "non violence declaration," adding that they are receiving "non-violent resistance" training by a special trainer who is sailing with them.
"We do not hit, or yell. If IDF soldiers board the ship and try to prevent it from reaching Gaza, we will try to protect our vessel with our bodies," he said.
The Estelle set off originally from Sweden and arrived in the Gulf of Naples on Thursday after a tour of Europe, including Finland, France and Spain.
A second attempt to break the blockade last summer with a coalition of ships setting sail from Athens failed after Greek authorities intercepted the boats, banning their departure and towing back into port those who tried to leave.
In November last year, the Israeli navy also intercepted two Irish and Canadian ships carrying 27 passengers and crew that were trying to breach the blockade in international waters off Gaza.
Shahar Chai contributed to the report