Dynamic cultivation of human mesenchymal stem cells in a rotating bed bioreactor system based on the Z RP platform.
Because the regeneration of large bone defects is limited by quantitative restrictions and risks of infections, the development of bioartificial bone substitutes is of great importance. To obtain a three-dimensional functional tissue-like graft, static cultivation is inexpedient due to limitations in cell density, nutrition and oxygen support. Dynamic cultivation in a bioreactor system can overcome these restrictions and furthermore provide the possibility to control the environment with regard to pH, oxygen content, and temperature. In this study, a three-dimensional bone construct was engineered by the use of dynamic bioreactor technology. Human adipose tissue derived mesenchymal stem cells were cultivated on a macroporous zirconium dioxide based ceramic disc called Sponceram. Furthermore, hydroxyapatite coated Sponceram was used. The cells were cultivated under dynamic conditions and compared with statically cultivated cells. The differentiation into osteoblasts was initiated by osteogenic supplements. Cellular proliferation during static and dynamic cultivation was compared measuring glucose and lactate concentration. The differentiation process was analysed determining AP-expression and using different specific staining methods. Our results demonstrate much higher proliferation rates during dynamic conditions in the bioreactor system compared to static cultivation measured by glucose consumption and lactate production. Cell densities on the scaffolds indicated higher proliferation on native Sponceram compared to hydroxyapatite coated Sponceram. With this study, we present an excellent method to enhance cellular proliferation and bone lineage specific growth of tissue like structures comprising fibrous (collagen) and globular (mineral) extracellular components.
Institut für Technische Chemie, Leibniz Universität Hannover, 30167 Hannover, Germany.
Solvig
S
Diederichs
Stefanie
S
Röker
Dana
D
Marten
Anja
A
Peterbauer
Thomas
T
Scheper
Martijn
M
van Griensven
Cornelia
C
Kasper
D000273:Adipose Tissue / Q000166:cytology; D000469:Alkaline Phosphatase / Q000378:metabolism; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002454:Cell Differentiation; D004396:Coloring Agents; D005947:Glucose / Q000378:metabolism; D006801:Humans; D019344:Lactic Acid / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000166:cytology; D016062:Porosity; D013050:Spectrometry, Fluorescence; D023822:Tissue Engineering / Q000295:instrumentation* / Q000379:methods; D054457:Tissue Scaffolds; D015040:Zirconium / Q000737:chemistry*
D004396:Coloring Agents; D019344:Lactic Acid; D015040:Zirconium; D000469:Alkaline Phosphatase; D005947:Glucose; C028541:zirconium oxide