Interprofessional collaboration (IPC) has been shown to improve patient outcomes, cost efficiency, and health professional satisfaction, and enhance healthy workplaces. We determined the attitudes of primary care physicians and nurses towards IPC and factors facilitating IPC using a cross-sectional study design in Singapore. A self-administered anonymous questionnaire, based on the Jefferson Scale of Attitudes toward Physician-Nurse Collaboration (JSAPNC), was distributed to primary healthcare physicians and nurses working in National Healthcare Group Polyclinics (N = 455). We found that the mean JSAPNC score for physicians was poorer than that for nurses (50.39 [SD = 4.67] vs. 51.61 [SD = 4.19], respectively, mean difference, MD = 1.22, CI = 0.35-2.09, p = .006). Nurses with advanced education had better mean JSAPNC score than nurses with basic education (52.28 [SD = 4.22] vs. 51.12 [SD = 4.11], respectively, MD = 1.16, CI = 0.12-2.20, p = .029). Male participants had poorer mean JSAPNC score compared to females (50.27 [SD = 5.02] vs. 51.38 [SD = 4.22], respectively MD = 1.11, CI = 0.07-2.14, p = .036). With regression analysis, only educational qualification among nurses was independently and positively associated with JSAPNC scores (p = .018). In conclusion, primary care nurses in Singapore had more positive attitudes towards IPC than physicians. Among nurses, those with advanced education had more positive attitudes than those with basic education. Greater emphasis on IPC education in training of physicians and nurses could help improve attitudes further.