| Summary: | 碩士 === 中原大學 === 醫學工程研究所 === 97 === The proliferation and differentiation of osteoblasts are important event
during bone turnover and are controlled both by local growth factors as well as
by systemic hormones. In their final phase of differentiation, osteoblasts become
embedded deep within the mineralized bone matrix during bone formation and
become osteocytes. Despite it is well known that the embryonic stem cells
(ESCs) are totipotent which can differentiate into a variety cell types, the
mechanism of stem cells osteogenesis in a multi-cellular environment is still
unknown. To further understand the interaction between ESCs and osteoblasts
during the osteogenesis, in this study, at first, we were interested in the
morphology of mouse embryonic stem cell line – C3H10T1/2 during the
osteogenic differentiation, cocultured with mouse MC3T3-E1 cell line in this
direct coculture model. Secondly, Different ratios of conditioned medium
collected from MC3T3-E1 were additional added into the medium for murine
C3H10T1/2 cell culture in this indirect coculture model. The osteoblast may
play a role on inducing or regulating the ESCs osteogenesis.
In the direct coculture system, direct coculture with different density
MC3T3-E1 cells and C3H10T1/2 resulted in increased cell number and alkaline
phosphatase activity expression, and measured by increased of deposited
calcium. This coculture system appears to enhance osteoblastic differentiateon
and produce extracellular mineralizing matrix.In the indirect coculture system,
there was a significant increase in cell number and alkaline phosphatase activity
expression in the presence of MC3T3-E1 CM in C3H10T1/2 cell. But the
amounts of deposited calcium were low. We propose MC3T2-E1 CM may
support initial proliferation and ALP activity but do not alter the ability of the
osteoblasts to produce extracellular mineralizing matrix.
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