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Just as with the preceding decade, the early 1620s saw ongoing upheaval in Artemisia Gentileschi's life. Her son Cristofano died. Just as she arrived in Rome, her father Orazio departed for Genoa. Immediate contact with her lover Maringhi appeared to have lessened. By 1623, any mention of her husband disappears from any surviving documentation.

Her arrival in Rome offered the opportunity to cooperate with other painters and to seek patronage from the wide network of art collectors in the city, opportunities that Gentileschi grasped. One art historian noted of the period, "Artemisia's Roman career quickly took off, the money problems eased". Large-scale papal commissions were largely off-limits, however. The long papacy of Urban VIII showed a preference for large-scale decorative works and altarpieces, typified by the baroque style of Pietro da Cortona. Gentileschi's training in easel paintings, and perhaps the suspicion that women painters did not have the energy to carry out large-scale painting cycles, meant that the ambitious patrons within Urban's circle commissioned other artists.

But Rome hosted a wide range of patrons. Fernando Afan de Ribera, 3rd Duke of Alcala, a Spanish nobleman, acquired her Penitent Magdalene, Christ Blessing the Children, and David with a Harp. During the same period she became associated with Cassiano dal Pozzo, a humanist and a collector and lover of arts. Dal Pozzo helped to forge relationships with other artists and patrons. Her reputation grew. The visiting French artist Pierre Dumonstier II produced a black and red chalk drawing of her right hand in 1625.

The variety of patrons in Rome also meant a variety of styles. Caravaggio's style remained highly influential and converted many painters to following his style (the so-called Caravaggisti), such as Carlo Saraceni (who returned to Venice in 1620), Bartolomeo Manfredi, and Simon Vouet. Gentileschi and Vouet would go on to have a professional relationship and would influence each other's styles. Vouet would go on to complete a portrait of Artemisia. Gentileschi also interacted with the Bentveughels group of Flemish and Dutch painters living in Rome. The Bolognese School (particularly during the 1621 to 1623 period of Gregory XV) also began to grow in popularity, and her Susanna and the Elders (1622) often is associated with the style introduced by Guercino.

Although it is sometimes difficult to date her paintings, it is possible to assign certain works by Gentileschi to these years, such as Portrait of a Gonfaloniere, today in Bologna (a rare example of her capacity as portrait painter), and Judith and her Maidservant, today in the Detroit Institute of Arts. The Detroit painting is notable for her mastery of chiaroscuro and tenebrism (the effects of extreme lights and darks), techniques for which Gerrit van Honthorst and many others in Rome were famous.