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Introduction It is estimated that mores nurses are needed in the future, nursing students’ career preferences are important to contribute the shortage of nursing manpower in Macao. Little is known regarding the preferences for nursing students’ career preferences and related factors among nursing students. Objective To explore and investigate the career preferences and related factors among nursing students in Macao. Methods A descriptive, cross-sectional survey study design was adopted to assess career preferences and related factors among nursing students in Macao. Results In total, 217 nursing students completed surveys. The most favorite career preferences among nursing students were community health nursing, paediatric nursing, medical surgical nursing, and obstetric & gynecological nursing; while the least career preferences among nursing students were oncology and hospice nursing, mental health nursing, and ER nursing. Factors were related to “gender”, “education”, and “nursing is my first study preference” among Macao nursing students for career preferences. Conclusion The senior nursing students prefer working in community health nursing and the junior and female nursing students prefer working in pediatric nursing and obstetric and gynecological nursing. The more understandings of nursing students’ career preferences, the better nursing curriculum design and implication to promote nursing students’ intention for working in the least career preferences work.