Effect of cryopreservation on some biological properties of rabbit adipose-derived mesenchymal stem cells / 中国实验动物学报

ABSTRACT

Objective To study the effect of cryopreservation on some biological properties of rabbit adipose -de-rived mesenchymal stem cells (rADMSCs).Methods rADMSCs culture was isolated by tissue explants adherent meth-od.Morphology of the primary cells was observed by inverted microscopy .Immunophenotypes of the rADMSCs were deter-mined using flow cytometry .The third passage cells were preserved in liquid nitrogen for 6 months, and then were thawed , and subcultured to passage 7.The growth curves of the cryopreserved cells were analyzed by MTT assay , and the cryopre-served cells were cultured in adipogenic and osteogenic medium , with non-cryopreserved rADMSCs as a control group .The adipogenic and osteogenic abilities of the rADMSCs were evaluated by oil red O staining , alizarin red staining and alkaline phosphatase activity assay , respectively.Results The rADMSCs cultured in vitro exhibited a spindle-shaped appearance and rapid growth expansion .Flow cytometry analysis revealed that the third passage rADMSCs were CD 44-and CD90-posi-tive, but negative for hematopoietic cells surface marker CD 45.The growth curves of cells in the experimental and control groups were “S” shaped, showing a non-significant difference between the two groups (P>0.05).The oil red O staining and alizarin red staining results were positive at 2 weeks after adipogenic and osteogenic induction .The ALP activities of the two groups were increased with osteogenic induction time , with a non-significant difference (P>0.05).Conclusions Cryopreservation does not affect the growth and differentiation pluripotency of rADMSCs significantly .