Improvement of in vitro culture method for umbilical artery smooth muscle cells

AIM:To establish and optimize the method for in vitro culture of human umbilical arterial smooth muscle cells (SMCs). METHODS: We counted each hole cell number in six orifices and statistically analyzed the number of primitive cells within the same time. Human umbilical artery vascular SMCs were cultured in vitro using optimized tissue-pieces inoculation. Alpha smooth muscle actin (alpha-SMA) and CD31 immunofluorescence staining methods and inverted phase contrast microscope were used for cell identification. RESULTS: Compared with the application of traditional cell culture technique, using the optimized cell cultivation this method to extract primitive cells was able to obtain more SMCs at the same time. The primary generation and subculture of SMCs grew well, and after frozen storage and recovery, the typical long spindle “peak-valley” growth of artery vascular SMCs could still be seen in the cultured cells. With subculture to the seventh generation, cell growth characteristics were stable and the cultured cells were confirmed as SMCs by alpha-SMA and CD31 immunofluorescence staining tests. CONCLUSION: Optimization of the tissue-pieces inoculation shortens the culture cycle time of human umbilical artery SMCs. Cultured cells have stable biological characteristics.