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The biological characteristics of microencapsulated human endostatin/293 cells and their inhibitory effects on the proliferation of human umbilical vein endothelial cells

Shanjun Cai, Wen-fang Zhan

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Abstract

Objective To observe biological characteristics of microencapsulated human endostatin/293 (hES/293) cells at different density and their inhibitory effects on the proliferation of human umbilical vein endothelial cells (HUVEC). Methods The microencapsulated hES/293 cells at different cellular density of 1 × 104 (group A) , 1 × 106 (group B) and 1 × 108 (group C) cells/ml were made by polyelectrolyte complexometry technology. The empty microcapsules were set as control group (group D). Each group has 6 samples. After 1, 3, 7, 14 and 35 days in culture, the number of total cells, viable cells was counted by trypan blue staining, and the survival fraction was measured. The grow status of hES/293 cells was measured by MTT assay, and the concentration of endostatin protein in supernatant was measured by enzyme linked immunosorbent assay (ELISA). HUVECs were co-cultured with hES/293 cells of group A,B and C. The proliferation of HUVEC at the 24, 72 and 120 hours after co-culture was measured by MTT assay. Results The number of total cells and viable cells were increasing and the survival fraction reached its peak after 3 days in culture in group A, B and C. The growth rate in group A was higher than that in group B and C after 3 days in culture (P<0.05), but the growth rate in group B was higher than that in group A and C after 7, 14 and 35 days in culture (P<0.05). The concentration of endostatin protein in the supernatant was the same in group A, B and C after 1 and 14 days in culture (P>0.05). However, group A had higher endostatin than group B and C after 3 days in culture, group B had higher endostatin higher than group A and C after 7 and 35 days in culture (P<0.05). The hES/293 cells of group A, B and C had no effects on the proliferation of HUVEC (P>0.05) after 24 hours co-culture, but can inhibit the proliferation of HUVEC after 72 or 120 hours co-culture (P<0.05). Conclusions The microencapsulated hES/293 cells at a density of 1 X 106 cells/ml can grow and survive, and release endostatin protein stably.The microencapsulated hES/293 cells at different density all can inhibit the proliferation of HUVEC. Key words: Endostatins/drug effects;  Endothelial cells;  Cells, cultured/ drug effects

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Objective To observe biological characteristics of microencapsulated human endostatin/293 (hES/293) cells at different density and their inhibitory effects on the proliferation of human umbilical vein endothelial cells (HUVEC). Methods The microencapsulated hES/293 cells at different cellular density of 1 × 104 (group A) , 1 × 106 (group B) and 1 × 108 (group C) cells/ml were made by polyelectrolyte complexometry technology. The empty microcapsules were set as control group (group D). Each group has 6 samples. After 1, 3, 7, 14 and 35 days in culture, the number of total cells, viable cells was counted by trypan blue staining, and the survival fraction was measured. The grow status of hES/293 cells was measured by MTT assay, and the concentration of endostatin protein in supernatant was measured by enzyme linked immunosorbent assay (ELISA). HUVECs were co-cultured with hES/293 cells of group A,B and C. The proliferation of HUVEC at the 24, 72 and 120 hours after co-culture was measured by MTT assay. Results The number of total cells and viable cells were increasing and the survival fraction reached its peak after 3 days in culture in group A, B and C. The growth rate in group A was higher than that in group B and C after 3 days in culture (P<0.05), but the growth rate in group B was higher than that in group A and C after 7, 14 and 35 days in culture (P<0.05). The concentration of endostatin protein in the supernatant was the same in group A, B and C after 1 and 14 days in culture (P>0.05). However, group A had higher endostatin than group B and C after 3 days in culture, group B had higher endostatin higher than group A and C after 7 and 35 days in culture (P<0.05). The hES/293 cells of group A, B and C had no effects on the proliferation of HUVEC (P>0.05) after 24 hours co-culture, but can inhibit the proliferation of HUVEC after 72 or 120 hours co-culture (P<0.05). Conclusions The microencapsulated hES/293 cells at a density of 1 X 106 cells/ml can grow and survive, and release endostatin protein stably.The microencapsulated hES/293 cells at different density all can inhibit the proliferation of HUVEC. Key words: Endostatins/drug effects;  Endothelial cells;  Cells, cultured/ drug effects

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Available abstract

Objective To observe biological characteristics of microencapsulated human endostatin/293 (hES/293) cells at different density and their inhibitory effects on the proliferation of human umbilical vein endothelial cells (HUVEC). Methods The microencapsulated hES/293 cells at different cellular density of 1 × 104 (group A) , 1 × 106 (group B) and 1 × 108 (group C) cells/ml were made by polyelectrolyte complexometry technology. The empty microcapsules were set as control group (group D). Each group has 6 samples. After 1, 3, 7, 14 and 35 days in culture, the number of total cells, viable cells was counted by trypan blue staining, and the survival fraction was measured. The grow status of hES/293 cells was measured by MTT assay, and the concentration of endostatin protein in supernatant was measured by enzyme linked immunosorbent assay (ELISA). HUVECs were co-cultured with hES/293 cells of group A,B and C. The proliferation of HUVEC at the 24, 72 and 120 hours after co-culture was measured by MTT assay. Results The number of total cells and viable cells were increasing and the survival fraction reached its peak after 3 days in culture in group A, B and C. The growth rate in group A was higher than that in group B and C after 3 days in culture (P<0.05), but the growth rate in group B was higher than that in group A and C after 7, 14 and 35 days in culture (P<0.05). The concentration of endostatin protein in the supernatant was the same in group A, B and C after 1 and 14 days in culture (P>0.05). However, group A had higher endostatin than group B and C after 3 days in culture, group B had higher endostatin higher than group A and C after 7 and 35 days in culture (P<0.05). The hES/293 cells of group A, B and C had no effects on the proliferation of HUVEC (P>0.05) after 24 hours co-culture, but can inhibit the proliferation of HUVEC after 72 or 120 hours co-culture (P<0.05). Conclusions The microencapsulated hES/293 cells at a density of 1 X 106 cells/ml can grow and survive, and release endostatin protein stably.The microencapsulated hES/293 cells at different density all can inhibit the proliferation of HUVEC. Key words: Endostatins/drug effects;  Endothelial cells;  Cells, cultured/ drug effects

Key concepts: Endostatin, Umbilical vein, Trypan blue, MTT assay, Molecular biology, Group A, Andrology, Cell culture

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