[Proliferation and differentiation of human osteoblasts from the nasal septum in a new perfusion culture system].
BACKGROUND
Today, perfusion culture systems are mainly used to investigate cellular physiology and to cultivate three-dimensional tissue complexes. As a rule, these systems are relatively expensive and do not enable continuous microscopic monitoring of the growing cells. Simple and inexpensive perfusion culture systems have not been available up to now.


METHODS
A novel perfusion culture system was developed in which the modular components consist of a mounting apparatus for inserting various media supply systems, microdispenser pumps, and laminar-flow culture chambers, each with a culture volume of 8 cm(3). The perfusion chambers were inoculated with human osteoblast cells from the tissue culture (5,000/cm(2)) and were perfused for 10 days after adherence of the cells (0.5 ml/min). As a control group, osteoblast-like cells were cultured in identically constructed culture chambers without medium perfusion. After 10 days, the cell counts were determined in accordance with the Coulter principle. Alkaline phosphatase was measured photometrically as a characteristic for differentiation.


RESULTS
Compared with the control group, three to four times the quantity of cells were produced within 10 days in the perfusion cultures. The alkaline phosphatase values were equally high or only slightly lower, indicating that osteoblast differentiation of the cells was maintained with a higher proliferation.


CONCLUSIONS
As large a number of in vitro proliferated cells as possible is a prerequisite for clinical application of tissue engineering. By continuously supplying medium, the tested perfusion culture system enables a higher rate of proliferation of osteoblast-like cells with maintenance of differentiation. Continuous microscopic monitoring of the cultures is possible using commercially available Petri dishes.
Bücheler
M
M
Klinik für HNO-Heilkunde/-Chirurgie, Universitätsklinikum, Sigmund-Freud-Strasse 25, 53105 Bonn, Deutschland. markus.buecheler@ukb.uni-bonn.de
Bücheler
B-M
BM
Hagenau
K
K
Hanke
G
G
Bootz
F
F
D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D044085:Microfluidics / Q000295:instrumentation*; D009300:Nasal Septum / Q000166:cytology*; D010006:Osteoblasts / Q000166:cytology*; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods