This research aimed to examine the contributions of different sources in developing high school chemistry teachers' practical knowledge of teaching with practical work in China. The examination was conducted on a theoretical framework in which the conceptualization of PCK and nine sources of teachers' PCK were involved. Through a questionnaire survey of 323 chemistry teachers, the findings included the following aspects. Firstly, among the nine sources, overall, 'teaching practices', 'curriculum materials', 'classroom observations', and 'peer coaching' were recognized as the core sources, whereas 'learning experience' was regarded as the peripheral source. Secondly, for the five components of PCK of teaching with practical work, the contributions of the nine sources are different. Thirdly, three sources were found to be significantly different among teachers who have different years of teaching experience: 'pre-service training experience', 'classroom observations', and 'peer coaching'. In the last part of this paper, the implications of the findings and the suggestions for the further studies were discussed.