Objective To investigate the cellular components and differentiation potential of cells in rabbit pericardial fluid, and to provide morphological basis for basic research and clinical application of pericardial cells. Methods Thirty adult New Zealand white rabbits, after aseptic thoracotomy, the pericardial fluid mixture was extracted, the fluid cells were centrifuged, isolated and cultured.The pericardial cellular morphology in the different generations was observed under the inverted microscope（The immunofluorescence staining method was used in the present study in order to analyze the pericardial cells phenotypes）.Their immunological phenotypes were analyzed by using immunofluorescence staining and the CD44, vimentin, CD45 and the number of cells positively expressed in the third generation cells were observed.The expression of CD44 and vimentin related molecules was detected by PCR. Results There was the cellular population with uniform morphology in the adult rabbit pericardial fluid. The cells with immunofluorescence positive staining for CD44 and vimentin were found in the pericardial fluid of rabbit, in addition, these cells possessed the immunofluorescence negative staining for CD45.After induction, they can differentiate into osteoblasts and adipocytes. Conclusion Rabbit pericardial fluid contains cells with multiple differentiation potentials, which may be of positive significance for myocardial repair.