Experimental Study On Co-culture Of Schwann Cells And Vascular Endothelial Cells In Different Proportions

Objective: Schwann cells and vascular endothelial cells were obtained for primary culture,and the fourth-generation SCs and VECs were cultured in different proportions in vitro.Observe the results of the cultivation in the experiment and find the optimal ratio of cultivation to provide an experimental basis for constructing vascularized tissue engineering nerves in vivo.Methods: Combine the co-culture of SCs and VECs in the medium at a ratio of 1: 1,1: 4,4: 1,8: 1,1: 8,and divide them into control groups according to the above ratio(ie group A—SCs alone culture is set Control group)and five experimental groups of B,C,D,E,and F.On days 1,3,5,and 7 of the co-culture of the two cells,SCs were counted and their growth curves were drawn to observe the effect of VECs on SCs.Results: 1.Schwann cell culture results show:The number of Schwann cells gradually increased with the increase of the culture time.By the 7th day,the cells were basically at the plateau stage.After the cells had been filled with the culture flask,they were passaged by trypsin digestion and identified as SCs by immunocytochemical staining with S-100? antibody.It is relatively uniform,with protuberances and multiple spindles,which can be seen to grow closely.2.Endothelial cell culture results show:VECs are monolayer flat round or strip-shaped fuselages at first,do not overlap with each other,the boundaries are clearer,they are polygonal,and they gradually converge and merge closely together.Basically cover the bottom of the bottle and confirm VECs with v WF and CD31 immunocytochemical staining.3.Direct co culture of Schwann cells and vascular endothelial cells:The number of a cells in the experimental group and the control group showed no significant difference(P > 0.05)on the first day of co culture.From the third day to the seventh day of co culture,the number of a cells in the experimental group B(1: l)and C(1: 4)increased And f(1:8)cell count was statistically different from that of control group A(P < 0.05),and there was no significant difference between experimental group D(4:1),e(8:1)and control group A(P > 0.05),but there was significant difference between experimental group D and experimental group E(P < 0.05).Conclusion: 1.The primary Schwann cells and vascular endothelial cells can be obtained through the steps of this experiment,which proves the feasibility of this subject and also provides a specific basis for the next co-culture experiment.2.The optimal ratio of Schwann cells and vascular endothelial cells to co-culture is 4:1,which provides a theoretical basis for the vascularization experiment of tissue engineering nerve.