Research Of The Differentiation Of BMSCs Into Adipocyte And The Inhibition Of Degrease Composition During The Differentiation

Objective: Harvest and separate adult human bone marrow mesenchymal stem cells(BMSCs) in vitro as described previously. Differentiate them into adipocytes. Study the influence of degrease composition on the adipogenic differentiation and research for a new therapy for patients with aplastic anemia(AA).Method:1. Observe thirty normal hospitalized patients'bone marrow which median age was 53.6 and twelve patients'with AA which median age was 56.4. Obtain their marrow by bone marrow puncture. Collect the Mononuclear cells(MNCs) layer. Inoculate them in culture flask at the density of 1×10⁶cells/ cm². Replace the media after five days and discard non-adherent cells. The experimental group: Collect the adherent cells of different passages in six-pore plaque at the density of 1×10⁵cells/ml. Incubate cells at 37℃in a humidified atmosphere of 5% CO₂ and fed by DMEM-LG medium replacement every four days. Change the media with adipogenic medium 24 hours later. Replace the adipogenic medium every three or four days. The control group: culture cells in high-glucose DMEM medium containing 10μg/ml insulin and 10% fetal bovine serum(FBS). Change medium every three or four days. Dilute collected MSCs with phosphate buffer saline(PBS) after 14 days. Dye cells with Red Oil O for five to ten minutes. Under light microscope count cells at random and the percentage of adipocytes. Determine the PPARγlevel of the two groups.2.We divide 30 patients'BMSCs into two groups at random. One was the inducing group in adipogenic medium, the other was the experimental...