"He (Putin) must now take responsibility vis-a-vis the rebels," Rutte told journalists in The Hague after pro-Russian separatists hindered access to the crash site where 298 died in Thursday's plane crash, 192 of them Dutch.
"The Netherlands and the world will see that he does what needs to be done," Rutte said.
"Given today's developments and the images from this morning, I sent a message to the president to once more exert his influence on the rebels," Rutte added.
"He (Putin) promised me his cooperation yesterday.
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"I told him that he must show the world that he wants to help."
Rutte said he was "shocked" by news images of "shameless" rebels handling passenger possessions and walking around the crash site.
Rutte said that he had spoken to Australian Prime Minister Tony Abbott and German Chancellor Angela Merkel and that they also said Putin must act.
"Unimpeded access and quick recuperation of the bodies is the number one priority," Rutte said.