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为获取足够数量活性良好的膀胱平滑肌种子细胞 ,本实验探索在旋转生物反应器内应用微载体技术快速扩增膀胱平滑肌细胞的方法。将培养的第二代新西兰兔膀胱平滑肌细胞应用Cytodex 3微载体在旋转生物反应器(RCCS)内进行动态培养 ,观测平滑肌细胞的生长情况和细胞代谢率 ,进行a 肌动蛋白免疫组化染色 ,并与常规方法培养平滑肌细胞进行比较。结果显示膀胱平滑肌细胞在Cytodex 3微载体上生长迅速、细胞代谢率高、生长倍增时间缩短。在培养第九天 ,细胞数量可达最初接种的 2 3倍。免疫组化显示平滑肌细胞活性良好。结果表明利用微载体细胞培养技术可简便快速地在体外扩增平滑肌细胞 ,可为构建组织工程化人造膀胱提供大量活性良好的平滑肌细胞
In order to obtain a sufficient number of well-activated bladder smooth muscle cells, this experiment explored the method of rapid expansion of bladder smooth muscle cells using microcarrier technology in a rotating bioreactor. The cultured second-generation New Zealand rabbit bladder smooth muscle cells were cultured in a rotating bioreactor (RCCS) with Cytodex 3 microcarrier to observe the growth of the smooth muscle cells and the cell metabolic rate, and then actin immunohistochemical staining. And compared with the conventional method to culture smooth muscle cells. The results showed that bladder smooth muscle cells grew rapidly on Cytodex 3 microcarriers with high cellular metabolic rate and shortened growth doubling time. On the 9th day of culture, the number of cells reached 23 times of the initial inoculation. Immunohistochemistry showed that smooth muscle cells were well activated. The results show that the use of microcarrier cell culture technology can be easily and quickly in vitro expansion of smooth muscle cells, for the construction of tissue-engineered bladder with a large number of well-activated smooth muscle cells