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.
Artigo
em Inglês
| WPRIM
| ID: wpr-249883
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>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Concentração Osmolar
/
Osteoblastos
/
Ratos Sprague-Dawley
/
Técnicas de Cultura de Células
/
Ácido Láctico
/
Reatores Biológicos
/
Meios de Cultura
/
Biologia Celular
/
Crescimento Celular
/
Glucose
Limite:
Animais
Idioma:
Inglês
Revista:
Biomedical and Environmental Sciences
Ano de publicação:
2007
Tipo de documento:
Artigo
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