The country's European Affairs minister, Sandro Gozi, warned anything said by Brussels officials could be manipulated and said the vote should be up to Scots.
His remarks, couched in diplomatic language, mark a clear departure from the stance of former European Commission President Jose Manuel Barroso, who said it would be difficult if not impossible for Scotland to rejoin the bloc.
Mr Gozi, a former EC official and diplomat, said: "We are not worried by the referendum in Scotland and we have no position on it because they are responsible for deciding their own future.
"We don't believe it is necessary for the European Union to take an official decision in advance and we don't want to have any kind of influence. We don't want to say anything because any word could be manipulated."
However, the centre-left Democratic Party MP, speaking to Barcelona newspaper La Vanguardia, added that the European Union was studying the implications of Scottish independence.
Italy holds the presidency throughout the second half of this year and its politicians would chair any meeting of EU leaders in the immediate aftermath of a Yes vote.
No senior Brussels official has been prepared to publicly support Mr Barroso's assertions on membership problems, though some believe a new Scottish member state may struggle to renew its share of the UK's budget rebate.
Mr Barroso has been replaced by former Luxembourg Prime Minister Jean-Claude Juncker, despite opposition to his appointment as commission president by UK Prime Minister David Cameron.
The SNP has said it can work with Mr Juncker, who is believed to be neutral on independence. First Minister Alex Salmond said the appointment had been a fiasco for Number 10, which put the UK "on the fast-track out of Europe".
Mr Salmond added: "The danger for us in Scotland is that we are dragged to the exit door too unless we take our destiny in our own hands this September.
"Cameron is playing a game of European roulette with Scotland's future, and with the jobs of many thousands of people in Scotland whose livelihoods depend on our links with the EU."
Meanwhile, Professor Sionaidh Douglas-Scott, a professor of European and human rights law at Oxford University, has added her voice to those legal experts who believe Scotland would retain EU membership post-independence.
She said: "Despite assertions to the contrary from UK lawyers, EU lawyers and EU officials, any future independent Scotland's EU membership should be assured, and its transition from EU membership qua part of the UK to EU membership qua independent Scotland relatively smooth and straightforward.
"It would take the form of an internal enlargement of the EU using the procedure for treaty amendment in Article 48. These arguments are made on the basis of EU law itself, which, it is argued, provide all the resources necessary to assure an independent Scotland's EU membership through EU treaty amendment, and not through a cumbersome accession process as a new member state."