Three-dimensional expansion: in suspension culture of SD rat's osteoblasts in a rotating wall vessel bioreactor / 生物医学与环境科学(英文)
Biomedical and Environmental Sciences
; (12): 91-98, 2007.
Article
in En
| WPRIM
| ID: wpr-249883
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To study large-scale expansion of SD (Sprague-Dawley) rat's osteoblasts in suspension culture in a rotating wall vessel bioreactor (RWVB).</p><p><b>METHODS</b>The bioreactor rotation speeds were adjusted in the range of 0 to 20 rpm, which could provide low shear on the microcarriers around 1 dyn/cm2. The cells were isolated via sequential digestions of neonatal (less than 3 days old) SD rat calvaria. After the primary culture and several passages, the cells were seeded onto the microcarriers and cultivated in T-flask, spinner flask and RWVB respectively. During the culture period, the cells were counted and observed under the inverted microscope for morphology every 12 h. After 7 days, the cells were evaluated with scanning electron microscope (SEM) for histological examination of the aggregates. Also, the hematoxylin-eosin (HE) staining and alkaline phosphatase (ALP) staining were performed. Moreover, von-Kossa staining and Alizarin Red S staining were carried out for mineralized nodule formation.</p><p><b>RESULTS</b>The results showed that in RWVB, the cells could be expanded by more than ten times and they presented better morphology and vitality and stronger ability to form bones.</p><p><b>CONCLUSIONS</b>The developed RWVB can provide the culture environment with a relatively low shear force and necessary three-dimensional (3D) interactions among cells and is suitable for osteopath expansion in vitro.</p>
Full text:
1
Index:
WPRIM
Main subject:
Osmolar Concentration
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Osteoblasts
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Rats, Sprague-Dawley
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Cell Culture Techniques
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Lactic Acid
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Bioreactors
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Culture Media
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Cell Biology
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Cell Enlargement
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Glucose
Limits:
Animals
Language:
En
Journal:
Biomedical and Environmental Sciences
Year:
2007
Type:
Article