[3D-Culturing of human osteoblastic cells with vessel like nutrient supply].
OBJECTIVE
The treatment of large, critical-size bone defects is a major therapeutic problem in orthopaedic and reconstructive surgery. The engineering of bone tissue could be used to replace lost bone mass. However, scaffolds seeded with vital cells and cultured in vitro suffer from poor oxygen and nutrient supply centrally, when the constructs exceed a critical volume. Therefore, we have established an osteoblastic cell culture in a new 3D-culture chamber with an artificial, vessel-like central membrane, allowing continuous nutrient supply.


METHODS
Human osteoblasts were cultured in a 3D-like manner using a perfusion chamber for one week. In this system, the nutrient supply is guaranteed by a vessel-like, semipermeable polysulfone membrane with a continuous flow of medium. After fixation and cryosectioning, histological and immunohistological staining and scanning electron microscopy was carried out.


RESULTS
Examinations reveal 3D cell growth around the central vessel. Formation of an extracellular matrix, rich in collagen type I and fibronectin, was detected immunohistochemically. Furthermore, we demonstrated cell adherence to the membrane and examined the surface morphology by scanning electron microscopy.


CONCLUSIONS
The innovative approach for 3D-culturing of human osteoblasts in a system with a central nutrient supply opens up new possibilities for the in vitro cultivation for tissue engineering.
Tischer
T
T
Chirurgische Klinik und Poliklinik Innenstadt der Ludwig-Maximilians Universität, München.
Schieker
M
M
Stengele
M
M
Pautke
C
C
Neth
P
P
Jochum
M
M
Mutschler
W
W
Milz
S
S
D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media / Q000378:metabolism; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005109:Extracellular Matrix / Q000502:physiology* / Q000648:ultrastructure*; D006801:Humans; D008567:Membranes, Artificial; D010006:Osteoblasts / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods
D003470:Culture Media; D008567:Membranes, Artificial