The lounge was in April found to be discriminatory by the state's civil and administrative tribunal after a complaint by a man refused entry to the exhibit.
The tribunal ruled MONA had 28 days to stop refusing entry to the lounge to people who didn't identify as ladies.
Artist and curator Kirsha Kaechele announced on Tuesday the gallery had lodged an appeal with the Supreme Court of Tasmania.
"It's worth exercising the argument, not only for the ladies lounge, but for the good of art and the law," she said.
"We need to challenge the law to consider a broader reading of its definitions as they apply to art ... as well as the right for conceptual art to make some people (men) uncomfortable.
"Ladies love the lounge - a space away from men - given what we have been through for the last several millennia, we need it."
Ms Kaechele said the tribunal took too narrow a view in terms of women's historical and ongoing societal disadvantage.
They did not recognise how the experience of the ladies lounge could promote equal opportunity, she said.
The lounge has been temporarily closed but there are plans for it to be reopened as a "church", "bathroom" or place of "education" under exemptions to discrimination laws.
Ms Kaechele said she could see circumstances in which men would be allowed in the lounge.
"We might allow them in on Sunday - and this would be part of our engagement (with an exemption) … as a school," she said.
"Perhaps women could bring their laundry in and every Sunday we could allow the men in for a few hours of instructions on folding and ironing."
Ms Kaechele thanked Sydney man Jason Lau for lodging the complaint, saying it took a "special person" with real bravery to pursue the action.
"He's taken heat but I think we should stop giving him a hard time," she said.
"Without Mr Lau, the artwork would really be not notable.
"(We) need to move on to the horribleness of men in general, rather than a single man."
Ms Kaechele said she would temporarily move Picasso works from the lounge to the women's toilets so they could be viewed without interruption.
The lounge housed some of the gallery's most important works, including a Sidney Nolan, and ladies were served by male butlers.
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