Effects of resveratrol on pig primary preadipocytes proliferation, differentiation and transcription expression of Sirt1 gene / 生物工程学报

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.

Three-dimensional spheroid model for cultivating WB-F344 cells in simulated microgravity / 生物工程学报

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.