[Isolation, culture and identification of mouse dental follicle cells and their heterogeneity].
OBJECTIVE
To establish a method for isolation and culturing mouse dental follicle cells and to identify its origin; meanwhile, the biological characteristics were detected.


METHODS
Mandibular first molars from 7-day-old Balb/c mice were digested with 1% trypsin; subsequently, the dental follicle was enucleated from the tooth germ and cultured. Vimentin and cytokeratin were examined to identify its origin. The shape of dental follicle cells were observed by HE staining. Gomori modified calcium-cobalt staining method was used to examine alkaline phosphatase (ALPase) activity of dental follicle cells. Immunocytochemistry was used to detect the expression of collagen type I and osteocalcin (OCN).


RESULTS
The isolated cells were pleomorphism and there were three basic types of cells: some were cuboidal/polygonal; some were elongated, spindle-shaped, fibroblast-like cells; a minor, third cell type was very thin and elongated. 2 or 4 nucleoli were within each nucleus.The cytoplasm of the first two cell types was filled with abundant granules and the cells had many filipodia. The expression of vimentin was positive while that of the cytokeratin was negative. Some but not all follicle cells expressed ALPase, collagen type I and OCN.


CONCLUSIONS
The cultured dental follicle cells were originated from mesenchyme, consisted of several cell phenotypes and were heterogeneous.
Ge
Shao-hua
SH
Shanghai Institute of Stomatology, Ninth People's Hospital, School of Stomatology, Shanghai Second Medical University, Shanghai 200011, China. geshaohua1973@163.com
Li
De-yi
DY
Yang
Pi-shan
PS
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D003795:Dental Sac / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008963:Molar / Q000166:cytology; D014746:Vimentin
D014746:Vimentin; D000469:Alkaline Phosphatase