Comparsion of cell-suspension and explant culture of mouse corneal epithelial cells in mice / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

AIM: To compare cell-suspension and explant culture of mouse corneal epithelial cells (MCEC).METHODS: MCEC were cultured by cell-suspension culture and explant culture, respectively. Colony forming efficiency (CFE) and cell proliferation were determined. The expression of corneal epithelial progenitor cell marker p63 and K19, as well as differentiation marker K12 was investigated by Western blotting.RESULTS: Twenty of 25 (80％) cornea explant were successfully subcultured to passage1 (P1), while only 12％ cell-suspension culture were successfully subcultured to P1. There were statistical significance between explant culture and cell-suspension culture (P<0.01). Up to 55％ of P1 cells in explant culture were passaged over P10 and were stably subcultured though at least 25 passages. However, cells cultured in suspension culture never achieved confluence in P2. CEF of P1 in explant culture was higher than P1 in cell-suspension culture (P=0.02) and CEF of P20 in explant culture was higher than P1 in explant (P=0.001). Immunostaining images showed expression of p63 and K19 in cell-suspension culture P1 and explant culture P1 and P20. K12 was expressed in P1 of both cell-suspension culture and explant culture, however, there was not K12 expressed in P20 of explant culture.CONCLUSION: In MECE culture, compared with cell-suspension culture, the explant culture is a preferable option.