Objectives  To establish a human fetal cardiomyocyte culture and to investigate whether the genes that encode transporters that may influence influx or efflux of bile acids are expressed in human fetal cardiomyocytes. Design  Laboratory study. Setting  Imperial College London. Sample  Six fetal hearts were obtained at the time of termination of pregnancy at 12–13 weeks of gestation and used to generate primary human cardiomyocyte cultures. Methods  To confirm the presence of cardiomyocytes, the cells were incubated with monoclonal antibodies to sarcomeric α‐actinin and anticardiac myosin heavy chain. Real‐time reverse transcription polymerase chain reaction was used to establish whether transcripts of genes that may influence bile acid transport are present in the culture (NTCP, BSEP, MDR3, FIC1, MRP2, MRP3, OATP‐A, OATP‐C, OATP‐D, OATP‐E) and whether taurocholate administration alters messenger RNA (mRNA) expression. Main outcome measures  Relative mRNA expression of genes of interest. Results  Real‐time polymerase chain reaction demonstrated the presence of mRNA for BSEP, MDR3, FIC1, OATP‐C, OATP‐D and OATP‐E in fetal heart. Four transcripts remained in the cardiomyocyte culture (BSEP, MDR3, FIC1 and OATP‐D), and we demonstrated the influence of taurocholate on gene expression. Conclusions  We have developed an in vitro model of the fetal heart that may be used for studies of the cardiac effect of endobiotics, e.g. bile acids, or of specific agents that may be used to treat the mother or fetus in pregnancy.