Vice President Mike Pence, chosen to lead the US delegation, stood as the American contingent of athletes entered the Olympic stadium, but according to the Associated Press, did not stand and applaud when athletes from the two Koreas walked together.
Cameras caught Pence not standing as the 143 athletes entered the stadium.
Speaking to reporters during a flight on Saturday, Mr Pence said: "There is no daylight between the United States, the Republic of Korea and Japan on the need to continue to isolate North Korea economically and diplomatically until they abandon their nuclear and ballistic missile programme".
The officials said it was fair to say both Pence and the North Koreans rebuffed South Korean President Moon Jae-in's efforts to facilitate a meeting between North Korea and the US.
Some in the arena were apparently taken aback by the North Korean cheerleaders - the special squad sent by the North Korean leader - who continued to sing and sway in unison while drowning out music performed by South Korean numbers on a nearby stage.
At a post-match press conference, North Korean player Jong Su Hyon sang the joint team's praises, saying, "One united is stronger than two divided".
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But North Korea's record on human rights and the growing threat from its nuclear weapons program appeared out of mind. North and South Korea teams entered together under the Korean Unification Flag during the Opening Ceremony.
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Kim Yo-jong's arrival was broadcast live on South Korean television. Now, in South Korea's second round hosting the worldwide sporting event, it seems the friction between the nations may drop to its lowest point in decades.
NBC will air the opening ceremony during its Friday primetime coverage starting at 8 pm ET / 5 pm PT.
Kim Yo Jong is probably Kim's closest confidant and is a senior cadre in North Korea's ruling party.
Abe shook hands and spoke with Kim Yong Nam at an Olympics reception, in his first conversation with a high-ranking North Korean official under Kim Jong Un.
He only stood up for the U.S. team, even though other dignitaries rose to their feet to applaud the joint arrival of athletes from both North and South Korea.
After being greeted at the airport by the head of South Korea's Unification Ministry, the delegation went from Seoul to the Olympic host city of Pyeongchang by high-speed train.
According to South Korea's Yonghap News Agency, internet-based telecasts were disrupted, and some people who bought tickets online for the opening ceremony said they couldn't print their tickets.