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There were 72 applications competing for a place in last year's Challenger Series, startups Paper Not Foil, Elixir and The Pet Bistro gaining a place in the programme.
Recently appointed Startup Dunedin chief starter Donna Hall said following the "outstanding success" of last year's programme, she wanted to scale things up.
"There's a lot of great content in the first month that's able to be shared across a higher number of startups, prior to us going into the one-on-one phase of the programme," she said.
Startup Dunedin would offer a month-long series of workshops for up to 10 applicants.
"At the end of the first month of workshops, three startups will be selected to complete an extended five-month programme," she said.
The three new businesses could each win up to $65,000 worth of services and a place in a five-month coaching programme featuring Deloitte, Gallaway Cook Allan, Startup Dunedin and a proven pool of local and national mentors.
Applications are open until Friday.
Startup Dunedin chairwoman Sarah Ramsay said the new process would allow more time to evaluate the final 10 and select the three for the programme. Startup Dunedin would ensure the remaining seven got the recognition and support they needed.
"Last year it was extremely tough for us to narrow down to just three startups," she said.
Mrs Ramsay said last year each of the three winners was paired with mentorship and in-kind sponsorship during a four-month process, which resulted in increased sales, international distribution agreements and improvements in business processes.
Shortlisted applicants will make a pitch on May 1 and 2, with the successful 10 doing a month of the programme, before making a second pitch on May 30 for the final three spots.
Mrs Hall said the programme sponsors Deloitte, Gallaway Cook Allan, Grow Dunedin Partnership, Enterprise Dunedin and Creative HQ had all renewed their commitment.
Enterprise Dunedin director John Christie said the Challenger Series specifically aimed to grow businesses which had the potential to upscale and contribute to Dunedin's future economy
"The start-up ecosystem has shown good signs of growth and activity over the last 12 months," Mr Christie said.
Last year's three businesses secured investment, customers and connections through the support of the Challenger Series, he said.
"This has set the foundation for success and we expect more startups will have an appetite to access this valuable programme, enabling them to establish and grow in the city," Mr Christie said.