Freedom of speech
Wilders, who said he was "angry and disappointed" with Wednesday's ruling, has defended his remarks were within the limits of freedom of speech.
"I know that I spoke the truth and didn't say anything punishable," he said, vowing to "fight like a lion".
The politician's 17-minute film, Fitna, was called "offensively anti-Islamic" by UN chief Ban Ki-moon and its screening in The Netherlands in 2008 prompted protests in much of the Muslim world.
Prosecutors had initially declined to charge Wilders, citing freedom of speech in dismissing dozens of complaints.
But an appeals court last January ordered prosecutors to put the MP on trial, saying politicians could not make "statements which create hate and grief".
Wilders has become one of the Netherland's leading politicians, with his Freedom Party emerging last year as the country's second-largest in the European Parliament.
Recent polls have also indicated the party also stands a chance to become the largest in the Dutch Parliament in national elections due in May 2011.