Effects of cryopreservation on biological characteristics of cultivated corneal limbal stem cell / 眼科

ABSTRACT

Objective To explore the effects of cryopreservation on biological characteristics of cultured limbal stem cells.Design Experimental study.Participants Twenty New Zealand white rabbits.Methods Twenty New Zealand white rabbits(40 eyes) were preparated of peripheral cornea with 2 mm and 2 mm circular shallow conjunctival limbal graft.Rabbit limbal stem cells(Rb-LSCs) used for cell-suspention culture were isolated form New Zealand white rabbit limbal tissues.The limbal stem cells,having formated monolayer cultured limbal stem cells,were cryopreservated one month,then thawing.By immunofluorescence staining,flow cytometry,inverted microscope,MTT assay,we compared the subculture of limbal stem cells cryopreservation before and after the structure and properties.Main Outcome Measurements Limbal stem cell activity and biological characteristics.Results Compared with freshing cells,the cells after cryopreservation in morphology were poor adherence late,small nuclear cytoplasm ratio,irregular shape.Im munofluorescence staining to detect the low-temperature frozen cells ABCG2 monoclonal antibody 81.7% positive rate decreased to 46.9%,the difference was statistically significant(P