The glossy advert depicts the lives of five "special" employees of Virgin Atlantic as they progress from childhood to their current jobs.
However, while the pilot is depicted as a confident, white, middle-aged male, the two females who feature are both immaculately dressed cabin staff with striking red lipstick and elaborately coiffured hair.
Teachers at the annual meeting of the Educational Institute of Scotland (EIS) in Perth said it was part of a wider culture of gender stereotyping.
"This advert tells you that if you are a boy you can be a pilot, but if you are a girl the best you can hope for is a job in the cabin, but only if you are beautiful and glamorous," said Mary Matheson, from the Aberdeen branch of the EIS.
"Most of the footage focuses on the attractive women which plays into the sexist imagery that a lot of young girls are exposed to today, that the body and the look is somehow more important than the substance of the person.
"I am surprised in today's society that an organisation as high profile as Virgin would put that out because it is reinforcing a stereotypical image that does not make young girls think differently about the sort of roles they can play in society."
The issue has been raised as part of wider concerns that female pupils are put off studying some subjects, such as science, because they are seen as leading to "male" careers.
There are continuing concerns over a shortage of female scientists and engineers, with just 36 female professors of physics across the UK in 2009-10 compared to more than 600 males.
And earlier this year, a study from the Organisation for Economic Co-operation and Development (OECD) concluded that girls lacked confidence in their ability in maths and science, and were therefore put off from applying for jobs in engineering and computing.
Delegates backed unanimously a motion calling on the ruling council of the EIS to investigate the gender imbalance in different subjects to find out which were worst hit and if any areas were bucking the national trend.
Globally, around 4000 of 130,000 airline pilots are women, according to the International Society of Women Airline Pilots. Fewer still are captains, with just 450 worldwide.
A spokeswoman for Virgin Atlantic acknowledged that only male pilots appeared in the advert, which was first aired last year.
But she added: "We do feature male and female crew in our adverts. The crew are an asset to Virgin Atlantic - they are one of the things that makes us stand out from our competitors.
"Part of that is the crew uniform, and the ladies' version being bright red is the iconic representation of Virgin Atlantic ... but it is certainly not intended to be sexist.
"In terms of our workforce, we do have female pilots, but a much higher percentage are male pilots - so it is reflective of the workforce."