"We need to immediately begin substantive talks... on questions of the political organisation of society and statehood for southeastern Ukraine with the goal of protecting the lawful interests of the people who live there," Putin was quoted as saying by Russian news agencies on a TV show broadcast in the far east of the country.
Russia has previously only called for greater rights under a decentralised federal system to be accorded to the eastern regions of Ukraine, where predominantly Russian-speakers live.
In the programme, taped on Friday, Putin did not directly address additional Western sanctions on Russia.
Putin, however, blamed the crisis in Ukraine on the West, accusing it of supporting a "coup" against pro-Kremlin president Viktor Yanukovych in February.
"They should have known that Russia cannot stand aside when people are being shot almost at point-blank," said Putin, adding that he did not have in mind "the Russian state but the Russian people."
Putin has denied that Moscow has sent regular troops to fight in Ukraine, but pro-Russian rebels have said that many Russian soldiers have volunteered while "on vacation".