Senator Hanson has been slammed as unfit to govern for the remarks and her party's painting of ex-SAS soldier-turned-Liberal frontbencher Andrew Hastie as a "traitor".
Prime Minister Anthony Albanese on Sunday said he was "stunned" to learn the One Nation leader criticised his mother for living in social housing for several decades.
Mr Albanese has publicly referred to his experience of growing up in public housing in inner-city Sydney with a single mother as being a motivation for his political career and vision as Labor leader.
"I'll make this point: if Pauline Hanson wants to criticise me, then that's fine," he told reporters in Sydney.
"But my mother was way out of Pauline Hanson's league, way out of it.
"I said during the last campaign, kindness isn't weakness. Tell you what's weakness: nastiness and spitefulness."
Senator Hanson made the remarks during an interview with Karl Stefanovic in February.
Trade Minister Don Farrell labelled the comments "appalling".
"I think Ms Hanson should apologise," the Labor frontbencher said.
"She wants dignity and respect from her fellow parliamentarians. Attacking the prime minister's mother like this is not a way to get respect and dignity from your colleagues."
She has also faced fierce criticism from Mr Hastie over a One Nation cartoon that branded him a traitor and on Sunday he joined his political opponents in denouncing her for the latest remarks.
The West Australian MP said Senator Hanson should "leave mums out of it", adding the comments about Mr Albanese's late mother were another sign she was unfit to lead.
He accused her of failing to take responsibility for her actions and hiding behind "childish" material in reference to the satirical cartoon.
"She's revealed in the last week that she doesn't have what it takes to be prime minister of this country," Mr Hastie told News24.
"When I put the pressure on her over the cartoon ... she showed herself to be rattled.
"If she were to be prime minister, is she going to run cartoons about all her political opponents? How is she going to devote her time?
"Is she serious? Does she have a worldview that we can actually understand? These are all legitimate questions for One Nation."
Senator Hanson has brushed off complaints about the "traitor" comments in the cartoon, saying it shouldn't be taken too seriously despite attracting criticism from across the political spectrum.
Her office was contacted for comment about the latest criticism.
Mr Hastie has been attacked by One Nation for testifying during the defamation trial of accused war criminal Ben Roberts-Smith, both of whom served in the elite Special Air Service Regiment in Afghanistan.
The Victoria Cross recipient has been charged over five alleged murders involving unarmed detainees during his deployment in Afghanistan between 2009 and 2012.
Roberts-Smith denies the allegations.
Displaying a photo of an Australian prisoner of war being beheaded by Japanese forces during the Second World War, Mr Hastie said Senator Hanson must clarify whether she believed it was acceptable to execute prisoners.
"She's got to come out and make her position very clear," he said.
"What's her standard for battlefield conduct?"
