A modified method to culture human osteoblasts from bone tissue specimens using fibrin glue.
BACKGROUND
To establish primary osteoblast cultures is a challenge. The methods for isolation mostly comprise digestion with extracellular matrix degrading enzymes after mincing the bone samples. These methods are labour intensive and lead to an inefficient recovery of cells. Therefore, the aim of this study was to develop a more reliable method for culturing human osteoblasts.


METHODS
Bone tissue specimens were obtained from 20 patients undergoing reconstructive operations. Bone specimens were dissected and put into petri dishes with the bottom covered with fibrin glue. To identify the nature of the outgrowing cells, cytological staining was performed, i.e. Von Kossa, Azan, Dahl's, alkaline phosphatase, and collagen type I.


RESULTS
Mean time interval of cellular outgrowth was 12 days after preparing the bone tissue specimens. Confluence of the cell cultures was reached after four to five weeks on average. All cells were positively stained using Von Kossa, alkaline Phosphatase and collagen type I. The matrix consisted of lime, calcium and collagens.


CONCLUSIONS
A simplified method to culture osteoblasts from all kinds of bone tissue specimens is presented. The fibrin glue allows firm adhesion of the specimens to the petri dish. This allows the cells to grow out without disturbance. Normally, due to movements during medium exchange the adhesive bonds are disrupted. The fibrin glue retains the adhesive bonds. This method allows studying human osteoblasts in different clinical settings.
van Griensven
Martijn
M
Department of Trauma Surgery, Hannover Medical School, Germany. Griensven.Martijn.van@MH-Hannover.DE
Zeichen
Johannes
J
Tschernig
Thomas
T
Seekamp
Andreas
A
Pape
Hans-Christoph
HC
D000368:Aged; D000469:Alkaline Phosphatase / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division / Q000187:drug effects; D024042:Collagen Type I / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D015718:Fibrin Tissue Adhesive / Q000494:pharmacology*; D006801:Humans; D008875:Middle Aged; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013194:Staining and Labeling; D014014:Tissue Adhesives / Q000494:pharmacology*
D024042:Collagen Type I; D015718:Fibrin Tissue Adhesive; D014014:Tissue Adhesives; D000469:Alkaline Phosphatase; D002118:Calcium