The Experiment Research Of Biological Characteristic Of HES/293 Cells Encapsulated In ACA Microcapsule And Inhibitory On Human Umbilical Vein Endothelial Cells

Objective The human Endostatin/293(hES/293) cells in the Alginate-Chitosan-Alginate-Alginate(ACA) microcapsules were made by polyelectrolyte com-plexometry technology. To study the growth and viability of human Endostatin/293(h-ES/293) cells in the Alginate-chitosan-Alginate(ACA) microcapsules of different cell density and the inhibitory effect on human umbilical vein endothelial cells (HUVEC), to compare the growth and viability between human Endostatin/293(hES/293) cells in the Alginate-Chitosan-Alginate(ACA) microcapsules and not. Methods Three groups were divided, group A,1X104 hES/293 cells per ml encapsulated in ACA microcapsu-le, group B with 1X106 hES/293 cells per ml and group C with 1X108 hES/293 cells per ml, each group included 6 samples. The hES/293 cells encapsulated in ACA micr-ocapsule were prepared by polyelectrolyte complexometry technology. The biological characters of hES/293 cells were observed by trypan blue staining.The cytoactive of cells in different cell density and the influences of human Endostatin/293 (hES/293) cells in the Alginate-chitosan-Alginate(ACA) microcapsules with different cell densit-y with different time(24-120h) on the HUVEC cells was detected by methyl thiazol-ye tetrazolium(MTT) and the albumen was detected by ELISA method. The biologic-al characters of hES/293 encapsulated and not were studied by flow cytometry respec-tively. Results hES/293 cells microcapsuled with different cell density were able to g-row and survivei n the microcapsules during observing days. The shape of microcaps-ules were sphericity, with smooth surface, hES/293 cells grew as single cell and scatt-ered in the microcapsules in the first 3 days. After the 7th day hES/293 cells began to assemble as cell clusters and scattered around the microcapsules. The number of total cells and living cells per capsule in different groups was increasing, the cell viability rate of all groups reached the top in the 3ed day.the cell viability rate of group B degraded gradually remaining stabilization in observing days, while the cell viability rate of group A and C reached the top until the 3ed day, and degraded strikingly after that day,reaching the lowest in the 35th day.By MTT method,The speed of cells in all groups reached the top in the 7th day and the speed of cells in group B keeped stabiliz-ation until the 35th day. The speed of cells of group A and C degraded strikingly after the 7th day and reached the lowest in the 35th day. By ELISA method, ES of group B and C were 217.63±27.96 ng/mL and 179.74±10.64 ng/mL in the 7th day,albumen contents in group B keeped stabilization until 35th day,while albumen contents in gro-up C degraded slightly. ES of group A was 175.37±26.23 ng/mL,degrading strikingly in 35th day. By the method of culturing hES/293 cells encapsulated in ACA and HUV-EC together,we conclude that in the first 24 hour HUVEC could grow without any in-fluence,four groups had notstatistic difference(p=0.536,0.620,0.151,0.901,0.397, 0.334),but the growth of HUVEC cells was significantly inhibited at 72 hour and 120 hour in the 3 experiment groups. At 72 hour,the growth of HUVEC cells was more significantly inhibited in group A, three groups had significant difference(p=0.022, p=0.006),while the growth of HUVEC cells was more significantly inhibited in group B at120 hour, three groups had significant difference(p=0.040,p=0.002). By MTT m-ethod, the cells microcapsuled grew slower than that of non-microcapsuled in the first 3 days, but both were similar in the fifth day. The OD-Value of two groups was notstatistic difference in 1,3,5,7days(t1=0.267,p1=0.795;t3=1.923,p3=0.083;t5=-1.363,p5=0.203;t7=1.424,p7=0.185).The percentage of S phase of non-microcapsule-d cells was higher than microcapsuled cells in the7th day, two groups had statistic diff-erence(t=2.725,p=0.034),and the difference lasted until the 14th day,two groups had statistic difference(t=2.545,p=0.044).Conclusions 1. This study showed that different cell density could influence cell cytoactive, hES/293 cells,1X106 cells per ml encapsulated in ACA microcapsule were able to grow and survive in the microcapsul-s,which biological characters and cytoactive were higher than that of 1X104 cells per ml and 1X108 cells per ml.2. hES/293 cells microcapsuled can release ES stably and inhibit the proliferation of HUVEC cells.3. The hES/293 cells encapsulated in ACA microcapsule have not only higher proliferating and metabolic activity, but also keep longer time than cells dissociational cultureed.