A modified method forin vitroisolation and cultivation of periosteal cells in rabbits / 中国组织工程研究
Chinese Journal of Tissue Engineering Research
; (53): 3523-3528, 2016.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-492686
Responsible library:
WPRO
ABSTRACT
BACKGROUND:
Periosteum is considered as a source of seed cels for cel therapydue toits biological features.OBJECTIVE:
To seek the optimal way to isolate and culture rabbit periosteal cels and identify their biological features.METHODS:
Rabbit periosteum on facies medialis tibiae was taken out under aseptic conditions. Periosteal cels isolated through the digestion of type II colagenase with the explants culture method were cultured in DMEM/F12 complete medium. Cel ultrastructure was observedunderan inverted microscope. Periosteal cel proliferation was determined bycel counting kit-8assay. Cel surface antigensCD90 and CD105 were determined using flow cytometry. Osteogenic andlipogenic induction mediums were applied to induce periosteal cels to differentiate into osteocytes and adipocytes, respectively. After 2 weeks of induction, cels were harvestedfor alizarin red staining and oil red O staining to assay the calciumnodules and lipid droplet. RESULTS ANDCONCLUSION:
The digestion of type II colagenase with the explants culture method shortened the period of primary cels culture and enhanced the survival rate, which causedhigher purity and stronger reproductive activity of harvested periosteal cels. Primary cultured periosteal cels grew in form of spindle spiral or paralel. Alizarin red andOil red O staining verified the multi-directional differentiation potentiality of periosteal cels. These findings suggest that the periosteal cels with high purity,strong reproductive activity,andmulti-directional differentiation potentialitycanbe harvested in short time using digestion of type II colagenase with the explants culture method.
Full text:
Available
Database:
WPRIM (Western Pacific)
Type of study:
Prognostic study
Language:
Chinese
Journal:
Chinese Journal of Tissue Engineering Research
Year:
2016
Document type:
Article