Research Of Three-dimensional Culture WB-F344Cell With Simulated Microgravity

Because of the limited donor liver, lacked effectual approach to proliferiation of hepatocytes in vitro and the deficient cryodepression condition, make the liver diseases therapy still faced many difficulties. Hepatic stem cells have the characteristic of bipotential differentiation that would be make it a good cell resource for liver diseases'cell therapy and tissue engineering. In this paper, to use Cytodex-3 as microcarrier, we established Three-dimensional (3D) culture of rat WB-F344 cells with rotary cell culture system (RCCS) to simulate microgravity environment. 3D culture of cells could closely mimic the in vivo situation with regard to superority than plane monolayer cultured cells. In order to study the rotating 3D culture model affection on cells proliferation and differentiation we examined cells morphology, proliferation, microstructure, mRNA expression of adhesion molecules,E-cadherin protein,AFP and albumin,polarization of membranes and cell transplantation and so on, aim to establish a potential way for clinic liver diseases'cell therapy and constructe a virtual model for tissue engineering .cells morphology,microstructure and proliferation were examined by optical microscope, transmission electron microscope, count the number of cells and flow cytometry method to test the variety of cells character under two culture conditions. The results demonstrated that cells were take on polyhedron with lots of micovilli and mitochondria, which grow well and packed together densely to form irregular aggregates in 3D culture condition, adjacent cells were connected with desmosome and tight junction. cells proliferation was increased compared with control, In the same phase, the percent of WB-F344 cell's cycle by flow cytometry demonstrated that G₀/G₁ and S phase increased, while G₂/M phase decreased(p<0.05).The membrane fluidity and the adhesive molecules expression of the WB-F344 cells were determined to research the variation between two conditions environment. The results demonstrated the polarization of membrane fluorescence and viscousness decrease in 3D condition compared with monolayer cultured cells, that's means the membrane fluidity increased(P﹤0.05). To detected mRNA expression of adhesive molecules, E-cadherin protein quantity by reverse ranscriptase-polymerase chain reaction (RT-PCR) and...