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6cells/ml in fresh media and passaged in T-25 flasks in an incubator. After two (2) passages, the cells were cultured directly in Tflasks and encapsulated medium by calcium alginate microcapsules, respectively. The cells were evaluated through live/dead assay, hematoxylin-eosin (HE) and alkaline phosphatase (ALP) staining. Moreover, Von-Kossa staining and Alizarin Red S staining were carried out for mineralized nodule formation. Following this, the oxygen consumption rates of osteoblasts in the earlier mentioned different cultures were measured on-line. The results showed that the osteoblasts performed well in its major functions after being examined by cellular morphology and viability, growth curve in growth kinetics, HE, ALP, Von-Kossa and Alizarin Red stains, which were capable of being used for further investigation as seed cells in bone tissue engineering. The osteoblasts cultured in static T-flask and encapsulated medium in vitro both proliferated well, and the oxygen consumption rates were 5.56 × 10 -6

Measurement of oxygen consumption rate of osteoblasts from Sprague-Dawley rat calvaria in different in vitro cultures