The Study On The Biological Characteristics Of HumanUmbilical Cord Blood-derived Mesenchymal Stem Cells And Differentiation Into Neurocyte-like Cells

Objective: To establish the method of in vitro cultur and expansion of human umbilical blood-derived mesenchymal stem cells (MSCs) and to investigate their biological characteristics.Be differentiated to nerve cell and neuroepithelial cell. Approach the possibility and mechanism from MSCs transplantation for the cerebral trauma.Method: 1 , Cell culture: We collect cord blood by the way of venepuncturing from cake uterine of common newborn. Mononuclearcell were separated under standered density gradient centrifugation.( Ficoll-Paque), cultivated in CO₂ incubator in which saturated humidity is 5% and temperature is 37.5 ℃. Culture fluid was changed regularly. 25 %trypsinase-0.01 %EDTA was used on serial subcultivation at the rate of 1:2 after the percentage of mixed cell reached to 80%. 2, Bionomics observation:To compare the growth character of MSCs from origin generation.,draw its growth curve.. Antigen sign of the amplificated MSCs was detected by flow cytometry;flow cytometer then compared to MSCs from medullary.All we do confirm that depurated mononuclearcel from cord blood is mesenchyme stem cell. 3, DMEM/F12 that contains 5%FBS, 10μ mol/Lretinoic acid, 10ng/mlbrain-derived neurotrophic factor and 10ng/mlciliary neurotrophic factor was added to amplification MSCs. NSE, NF-sign of nerve cell and GFAP- sign of glia cell could be detected through fluorescent staining and PT-PCR on guided MSCs. 4 , We put MSCs in animal model of craniocerebral injuried bandicoot,observ ing the transmutation of the sign of the nerve cell and the functional recovery of the headinjuried rat.