ABSTRACT
Three-dimensional (3D) culture of cells could closely mimic the in vivo situation with regard to cell function and microenvironment compared with plane monolayer cultured cells. In this paper, we established 3D culture of rat WB-F344 cells with rotary cell culture system (RCCS) to simulate microgravity environment, and examined cells proliferation, morphology, microstructure, E-cadherin protein quantity and mRNA expression of adhesion molecules by count the number of cells, optical microscope, transmission electron microscope and reverse transcriptase-polymerase chain reaction (RT-PCR). The results demonstrated that cells were polyhedron with lots of micovilli and mitochondria, which grow well and packed together densely to form irregular aggregates. Adjacent cells were connected with desmosome and tight junction. With the regard, the aggregates behaved 3D growth characteristics. Moreover, compared with control, mRNA level of Fibronectin and E-cadherin protein were increased, the changes maybe is the part mechanism in this microgravity simulated cells culture models which strengthened cells junction. This rotating 3D model might facilitate the study of interactions of cell-cell, cell-matrix and the mechanisms.
Subject(s)
Animals , Rats , Cadherins , Genetics , Cell Adhesion , Cell Culture Techniques , Methods , Cell Proliferation , Fibronectins , Genetics , Spheroids, Cellular , Weightlessness SimulationABSTRACT
1 approximately 3 days old Piglet's primary preadipocytes in vitro were cultured and treated with 0micromol/L (control group), 10microlmol/L (lower dose group), 20micromol/L(middle dose group) and 50micromol/L, 100micromol/L (higher dose group) RES. Cell proliferation and viability were analyzed by MTT assay. The degree of differentiation and adipogenesis were measured by Oil Red O staining extraction assay and the expression of Sirt1 (sirtuin) mRNA were detected by RT-PCR. The results showed the optical density (OD) of MTT and Oil Red O staining were all decreased, especially treated by 50micromol/L, 100micromol/L RES at 72h and 96h (P < 0.01); the ratio of OD of the expression of Sirt1 mRNA to that of beta-actin mRNA were increased after treated by 100micromol/L RES (P < 0.01). RES can inhibit proliferation and differentiation of pig preadipocytes in certain degree. Higher dose of RES can markedly decrease adipogenesis and prevent preadipocytes differentiation into adipocytes, which may be in part associated with its effect on increasing the expression of Sirt1 mRNA.
Subject(s)
Animals , Adipocytes , Cell Biology , Metabolism , Cell Differentiation , Cell Proliferation , Cells, Cultured , RNA, Messenger , Sirtuin 1 , Genetics , Stem Cells , Stilbenes , Pharmacology , Swine , Transcription, GeneticABSTRACT
<p><b>BACKGROUND</b>Internal metallic stents have been widely used in clinical practice, but a high postoperative restenosis rate limits its application. The purpose of this study was to determine the effect of intrabiliary radiation on muscle formation and biliary duct remodeling after biliary duct balloon injury in dogs.</p><p><b>METHODS</b>Twenty male dogs (15 - 20 kg) were randomly divided into treatment group (n = 10) and control group (n = 10). Balloon overstretching injury was induced using a balloon catheter placed across the biliary duct. Subsequently, a 103Pd radioactive stent was positioned at the target site in each animal in the treatment group, providing the injured biliary duct with a radiation dose of 12.58 x 10(7) Bq. Dogs in the control group received Ni-Ti stents. All the dogs were killed one month after initial injury. The injured sections were dissected free from the dogs, and were processed for histological and morphological study. Cross-sections were stained with hematoxylin-eosin, Masson's trichrome, and Verhoef-van Giesen. Muscle formation area and lumen area were determined using a computer-assisted image analysis system.</p><p><b>RESULTS</b>Compared with the control group, 103Pd radioactive stents significantly reduced muscle formation area (78.3%, P < 0.01), and percentage area of stenosis [control stents: (60.0 +/- 21.6)%, 103Pd radioactive stents: (31.6 +/- 9.5)%]. In addition, in the treatment group, the biliary duct lumen area was significantly larger than that in the control group (P < 0.01).</p><p><b>CONCLUSIONS</b>103Pd radioactive stents providing a radioactive dose of 12.58 x 10(7) Bq are effective in reducing muscle formation and biliary duct remodeling after balloon overstretching injury.</p>