Isolation, Culture And Differentiation Potential Of Rat And Rabbit Pericardial Cells In Vitro

BackgroundPericardial fluid(PF)is a clear slurry that exists between the wall layers of the serous pericardium.The pericardial fluid reduces the friction inside the pericardium by lubricating the epicardial surface and has a lubricating effect.Whether or not the pericardial fluid has cellular components has not been reported so far.ObjectiveTo understand the cellular components and differentiation potential of pericardial fluid in rats and rabbits,and to provide morphological basis for basic research and clinical application of pericardial cells.Methods1.The pericardial fluid was aseptically extracted,separated by Ficoll method,and centrifuged.Different cultured cells were taken and the morphology of the cells was observed under an inverted microscope.Immune coloration was performed on the pericardial cells by immunofluorescence technique,and the surface markers of pericardial cells were observed.2.The immunofluorescence technique was used to immunochromatize the rat and rabbit pericardial cells,and the expression of surface markers CD44,CD45,Vimentin,Sca-1 and nanog on the pericardial cells was observed.The primary antibody was replaced with PBS as a negative control.3.The third-generation rat and rabbit pericardial cells were induced into osteogenic and adipogenic cells,respectively,and the pericardial cells of the rats were induced into myocardium.Cells cultured in complete medium were used as controls.4.Total RNA was extracted from rabbit pericardial cells,and cDNA was synthesized by reverse transcription.Real-time quantitative PCR was used to detect the expression of CD44,CD45 and Vimentin.The difference of CD44 and Vimentin gene expression was detected by real-time PCR.GAPDH was used as an internal control.Results1.The rat and rabbit pericardial fluid contains a cell group with uniform morphology,good growth state and strong proliferative ability.The cell morphology is mostly polygonal or long fusiform,and the rat is more slender than the rabbit pericardial cell.Vortex-like growth,and rabbit pericardial cells more into an endothelial-like growth pattern..2.Immunofluorescence results showed that rabbit pericardial cells had positive expression of CD44 and Vimentin and did not express CD45.The results of PCR showed that the expression of Vimentin was significantly higher than that of CD44(p<0.05).The expression of CD29,Sca-1,CD44 and Vimentin was not expressed in rat pericardial fluid,and CD45 and nanog were not expressed.3.The third generation cells were taken for osteogenic,adipogenic or myocardial induction.PCR technology was used to detect the expression of related molecules and to analyze the difference in expression levels.ConclusionsRat and rabbit pericardial fluid contain multiple differentiation potential cells,which can differentiate into osteogenesis,adipogenic and cardiac differentiation.They may have positive implications for heart or pericardial repair..