ABSTRACT
<p><b>OBJECTIVE</b>To follow the fate of murine epidermal stem cells (ESCs) seeded in a syngeneic dermal equivalent implanted in vivo.</p><p><b>METHODS</b>Embryonic stem (ES) cells were induced in vitro to differentiate into ESCs. After stained with a fluorescent dye Hoechst 33342, these ESCs were seeded into a fibroblast-collagen-gelatin sponge complex, functioning as a dermal equivalent model, and implanted subcutaneously into 129/J mice, which were syngeneic to these stem cells. The fate of these cells was observed with HE staining, immunocytochemical staining or Van Gieson's staining.</p><p><b>RESULTS</b>These ESCs were clearly visible in the implant by fluorescent microscopy 3 weeks or longer after implantation. These cells remained viable, differentiated into hair follicle-like structure, glandular structure, and gave rise to additional structures displaying features resembling native dermis. A number of markers were expressed in the differentiated structures, including CD29 (integrin beta1 subunit) and cytokeratin 18 (CK18). No apparent rejection or severe side effects were observed at least during the 10 weeks following implantation.</p><p><b>CONCLUSION</b>Now that ESCs could survive in vivo in this dermal equivalent model, and differentiate into hair follicle-like structures as well as glandular structures, it is feasible to use these cells as seed cells in the study to fabricate dermal equivalent having the potential to develop dermal appendages.</p>
Subject(s)
Animals , Mice , Cell Culture Techniques , Cells, Cultured , Dermis , Cell Biology , Transplantation , Embryonic Stem Cells , Cell Biology , Epithelial Cells , Cell Biology , Mice, Inbred Strains , Skin Transplantation , Stem Cell Transplantation , Stem Cells , Cell Biology , Tissue EngineeringABSTRACT
Objective To establish a method for isolation and cultivation of human invasive pituitary adenoma-derived fibroblast in vitro and explore their biological properties in order to investigate the role of the fibroblasts in the invasive growth of the pituitary adenoma.Methods The fibroblasts were isolated from hu- man invasive pituitary adenoma tissue by different rate of adhesion.Primary culture and passage culture were carried out,and growth characteristics of fibroblasts were observed under phase-contrast microscope in primary and passage culture.Immunohistochemical technique was used to identify the fibroblasts.The cell growth curve was measured by MTT method.The ultrastructure was observed under electronmicroscope.Results The cultured invasive pituitary adenoma-derived fibroblasts showed active proliferative ability.The cell bodies was bigger than other tissue fibroblasts,and the cell shape was irregular.The rough endoplasmic reticulum, Golgi complex and ribosome were profuse in the cytoplasm.The positive expression rate of collagen[and vi- mentin of the fibroblasts were above 95%.Conclusion The method of different rate of adhesion is success- fully utilized to culture human invasive pituitary adenoma-derived fibroblasts.These cells showed active prolif- erative ability,and maybe have an importent function in the invasive growth of the pituitary adenoma.