The Comparation Of Multi-directional Differentiation In Fibroblasts From Multiple Organs In Neonatal Rats

BackgroundFibroblasts are abundantly distributed throughout connective tissues in the body and are very important in maintaining the structural and functional integrity.Fibroblasts have traditionally been viewed as a uniform cell type with supporting and secreting function regardless of the origins of tissue.Recent studies have proved that there are many subpopulations in fibroblasts,which have significant heterogeneity in cell morphology,growth characteristics,collagen synthesis,cytokines and surface receptors.More and more reports demonstrate fibroblasts and mesenchymal stem cell share much more in common than previously recognized.However,there is still lack of comparative studies in fibroblasts on the differences in the expression of molecular markers and differentiation capacity from different organs.ObjectivesThe aims of this study were to evaluate the differences in morphological characteristics,surface markers,and multiple differentiation potential of fibroblasts in four organs in neonatal rats heart,such as lung,kidney,liver and skin and to prove biology phenotypic differences,which would provide the direct expremential data for screening popular organs in organ repair.MethodsCombined enzymes digestion method was used to isolate and culture the fibroblasts derived from liver,lung,kidney and skin.Common molecular markers for fibroblasts such as vimentin,DDR2 and FSP1,stem cell markers nanog and c-kit were detected by RT-PCR,and immunofluorescence.The osteogenic and adipogenic differentiations of fibroblastswere performed by inductive culture in special mediums,and analyzed by Alizarin red and Oil red respectively.ResultsThe isolated fibroblasts had high purity and large volume by the method.There were obvious heterogeneity in enzyme digestion time.was different.Tissue blocks from lung,liver,kidney and skin were digested and the digestion time was 30 min,25 min,35 min and 80 min respectively.However,there are not obvious differente in cell morphology from multiple organs.DDR2 was partly expressed in the skin and lung.Interestingly,no expression of DDR2 was detected in liver and kidney.nanog expression in skin,liver and spleen had significant difference comparing with in lung(**P<0.01).Directed differentiation in vitro had demonstrated that skin fibroblasts had strong multiple differentiation potential.ConclusionsIn this study,we have demonstrated that there are heterogeneity in fibroblasts from four organs.DDR2 could not be used to the special molecular marker for fibroblasts from four organs,while combined expression of vimentin and FSP1 could be as effective molecular markers in fibroblast from four organs.Skin fibroblasts had the strongest multiple differentiation potential.