Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (10): 1547-1553.doi: 10.3969/j.issn.2095-4344.2014.10.011
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Wan Ying1, Ben Liang1, Guan Zi-qiu 2, Li Chao1, Zhang Shi-dong1, Nie De-zhi1
Online:
2014-03-05
Published:
2014-03-05
Contact:
Nie De-zhi, M.D., Technician in charge, TuoHua Biological Technology Company, Siping 136000, Jilin Province, China
About author:
Wan Ying, M.D., TuoHua Biological Technology Company, Siping 136000, Jilin Province, China
CLC Number:
Wan Ying, Ben Liang, Guan Zi-qiu, Li Chao, Zhang Shi-dong, Nie De-zhi. Effects of Corning® CellBIND® Surface medium on growth of human umbilical cord mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(10): 1547-1553.
Effects of Corning® CellBIND® Surface dish coated with poly-L-ornithine on cell adhesion Cell numbers and morphology were observed in different culture dishes within 24 hours as shown in Figure 1. Cells cultured in the Corning® culture dish were most adhered and spindle-shaped, resembling fibroblasts. Corning® CellBIND® Surface culture dish also exhibited the powerful effect on cell growth[15-22], and the number of adherent cells was increased with the time prolonged within 24 hours. Depending on the treatment of microwave plasma process for treating the culture surface, Corning® CellBIND® Surface culture dish coated with poly-L-ornithine exhibited a better effect on cell attachment than Corning® tissue culture polystyrene dish coated with poly-L-ornithine (Figure 2)."
Effects of Corning® CellBIND® Surface dish coated with poly-L-ornithine on cell proliferation Cells proliferating in the different culture surfaces were observed in 7 days as shown in Figure 3. The results showed the growth curve of hUMSCs presented inverted “S”, and cells exhibited logarithmic growth on days 3, 4, 5 and 6. Owing to the effect of poly-L-ornithine on cell adhesion, Corning® tissue culture polystyrene dish and Corning® CellBIND® Surface dish coated with poly-L-ornithine improved cell growth at 7 days compared with the culture dishes that coated without poly-L-ornithine. However, Corning® CellBIND® Surface dish exerted the better effect on cell proliferation under any treatment on the surface of dish compared with Corning® tissue culture polystyrene dish (Figure 3)."
Effects of Corning® CellBIND® Surface dish coated with poly-L-ornithine on cell adhesion and cell proliferative protein expressions To test the effect of Corning®CellBIND® Surface culture dish on cell attachment and cell proliferative protein expressions, first, we detected cell proliferative protein including cyclinA, cyclinE and PCNA expressions. The results showed that neither Corning® CellBIND® Surface culture dish, Corning® tissue culture polystyrene dish nor those coated with poly-L-ornithine influence the expressions of PCNA, cyclinA or cyclinE by western blot analysis (P > 0.05). Furthermore, the expressions of adhesion cytokines were examined and the expression of fibronectin was significant increased in Corning® tissue culture polystyrene dish coated with poly-L-ornithine and Corning® CellBIND® Surface culture dish coated with or without poly-L-ornithine (P < 0.05) (Figure 4)."
Effects of Corning® CellBIND® Surface dish coated with poly-L-ornithine on expression of cell surface markers We previously demonstrated that similarly processed hUMSCs expressed high levels of matrix receptors (CD73, CD90 and CD105) as assessed by flow cytometry. In addition, all adherent hUMSCs did not express CD45, CD34, CD14, CD19 or HLA-DR (data was not shown). In order to determine the effect of Corning® CellBIND® Surface culture dish on hUMSCs phenotype, we examined the levels of CD73, CD90 and CD105 in the four different groups. As our predicted, hUMSCs highly expressed CD73, CD90 and CD105 in the four groups as same as our isolated cells from human umbilical cords before. It implied that Corning® CellBIND® Surface culture dish coated with or without poly-L-ornithine did not affect the cell phenotype (Figure 5)."
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