The 58-year-old, who confirmed she had adopted the two four-year-old girls from Malawi on 8 February, captured them singing ‘Twinkle, Twinkle’ on the sofa while wearing matching striped pyjamas.
In the background, someone played the tune on the piano while they sang.
“A little night music,” Madonna captioned the photo on Sunday 19 February.
The video has had more than 500,000 views in just one day.
“Absolutely beautiful children,” one person commented on the video.
“Oh my gosh this is so sweet,” wrote another. “Look how happy they look. This is just wonderful. All the best wishes to you and your family.”
Madonna shared the first photo of the twins, Estere and Stelle, when she confirmed rumours that she was adopting.
Alongside the snap of her holding both of their hands, she wrote: “I can officially confirm I have completed the process of adopting twin sisters from Malawi and am overjoyed that they are now part of our family.
“I am deeply grateful to all those in Malawi who helped make this possible.”
Madonna shared another photo of her twins on 9 February encouraging others to donate or volunteer at their orphanage.
“There are 650 orphans at Home of Hope where Estere and Stelle have lived for four years since they were five days old,” she wrote.
“If you would like to get involved by volunteering or donations go to. Raising Malawi.org. Every little bit helps.”