Rohini Hattangadi joined the National School of Drama (NSD), New Delhi in 1971. She met Jayadev Hattangadi, who was also in the same batch as hers. Together, they received training under theatre Guru Ebrahim Alkazi. Upon graduation in 1974, Rohini was awarded the Best Actress award and also an award for the Best all-round student, while Jayadev who was training in direction, won the Best Director Award. Jayadev and Rohini married shortly thereafter. Rohini also underwent training in Indian classical dance forms, Kathakali and Bharatnatyam for more than eight years, under the guidance of Professor Surendra Wadgaonkar. Rohini started her career with Marathi stage. Once in Bombay from NSD, Jayadev and Rohini started a Marathi theatre group in Bombay, called ‘Awishkar’, which went on to produced over 150 plays.
Rohini Hattangadi got her first break with Saeed Akhtar Mirza's Arvind Desai Ki Ajeeb Dastaan in 1978. The film won the National Film Award for Best Film. She worked with him again in Albert Pinto Ko Gussa Kyon Ata Hai (1980). Her next film was Chakra (1981) by Rabindra Dharmaraj, starring Naseeruddin Shah and Smita Patil. Rohini’s performance in an international film Gandhi (1982), got her instant international recognition and won her many accolades. Mahesh Bhatt's Arth (1982) was a critically acclaimed film and she even received an award for her performance. Her performance in Govind Nihalani's Party (1984) also won her appreciation. In 1984, her films Mohan Joshi Hazir Ho! And Saaransh with Anupam Kher enabled to strengthen her position in the industry. Rohini did a few films like Insaaf Kaun Karega (1984), Bhavna (1984), Dharam Adhikari (1986), Jalwa (1987) and Andha Yudh (1987), all of which were average to poor. She worked with Govind Nihalani again in Aghaat (1985), and with Muzaffar Ali in Anjuman (1986) Her powerful yet small role in N. Chandra's Pratighaat (1987) gained her wide appreciation. Thikana (1987), Tamacha (1988), Shukriya (1988), Rukhsat (1988) were all flops, while Amitabh Bachchan-starrer Shahenshah (1988) was quite successful. She also appeared in the Naseeruddin Shah-starrer Hero Hiralal (1988). Rohini gave excellent comic performances in Pankaj Parashar's Chaalbaaz (1989) and in Ladaai (1989). She played Amitabh Bachchan’s mother in Agneepath (1990), where yet again she proved her mettle of delivering power-packed performances, though the film flopped. In 1991, Rohini worked with Girish Kasaravalli in Mane and Ek Ghar, and gave strong performances yet again. Benaam Badshah (1991) and Kurbaan (1991) were average. She also starred in the horror flick Raat in 1992. Rohini’s other hit films included Rajkumar Santoshi’s Damini – Lightning (1993) and Ghatak: Lethal (1996), and Rajkumar Hirani’s Munnabhai M.B.B.S. (2003). Krishna (1996), Gambler (1997), Qahar (1997), Rajkumar Santoshi’s Pukar (2000) did average business. Akele Hum Akele Tum (1995), Veergati (1995), Kabhi Na Kabhi (1998), Jaanam Samjha Karo (1999), Lajja (2001), unfortunately flopped. Rohini Hattangadi will be seen in the Himesh Reshammiya’s Karzzzz (2008).
Rohini and Jayadev have a son Aseem Hattangadi, who is also a theatre actor and acted in Badal Sircar’s play, 'Evam Indrajit', directed by Jayadev Hattangadi himself. In 1975 she won the ‘Best Actress Award' at the Maharashtra State Drama Festival, for her performance in ‘Changuna’ a Marathi adaptation of Federico García Lorca’s Spanish classic ‘Yerma’, produced by ‘Awishkar’, in Mumbai. She became the first woman to act in ‘Yakshagana’, a folk play from Karnataka titled 'Bhisma Vijay', directed by Dr. K. Shivaram Karanth, besides being the first woman in Asia to act in a Japanese Kabuki play, Ibaragi, directed of Shozo Sato, a well-known Japanese director. The play that stands out in her long and illustrious career is 'Aparajita', based on Bengali story by Nitin Sen, this 120 min. long, solo-act play, which displays her versatility. Rohini has performed in five plays directed by her husband including Medea, a Greek tragedy written by Euripides. She won the 2004 Sangeet Natak Akademi Award for her contribution to Indian theatre. Rohini’s work was also widely appreciated in Vijay Tendulkar’s ‘Mitra Chi Gosht’ directed by Vinay Apte, 'Hori’ based on Premchand’s ‘Godaan’, produced by IPTA, and most recently in Hindi play, ‘Kohra’, with Sudha Chandran, and Babul Bhavsar. Jayadev and Rohini Hattangadi, also run ‘Kalashray’, a centre for research, education in arts and talent encouragement in Mumbai, working with the underprivileged and developing tools for powerful communication. She has been seen in a number of TV serials including Mahayagya, Thoda Hai Thode Ki Zaroorat Hai, Teacher, etc.