OBJECTIVE: To establish the best primary and propagated culture approach of the human umbilical artery smooth muscle cells (HUASMC) and immortalized the primary cells. METHODS: The primary culture of HUASMC was performed using the primary-explant method, and the effects on the cells' growth characteristic by different culture mediums were conducted. Primary cultures of HUASMC derived from normal human umbilical arteries were transfected with recombination plasmid containing human telomerase reverse transcriptase (hTERT) open reading frame. The transfected HUASMC was analyzed for cell biological characteristics, and growth characteristics, and specific alpha-actin. RESULTS: Highly purified primary HUASMC were gained and could be propagated about 10 population doublings (PD). After PD 4, the cells lost the proliferation ability gradually. However, the transfected HUASMC could be propagated at least 20 population doublings and could express specific alpha-actin. CONCLUSION: The hTERT transfected HUASMC could keep the basic biological characteristic of smooth muscle cells and prolonged life span at least 20 PD. It might be a good cell model for the study of cardiovascular disease, myoma of uterus and chronic graft disfunction. It might also bring the hope to engineered vascularization in the future.
OBJECTIVE: To establish the best primary and propagated culture approach of the human umbilical artery smooth muscle cells (HUASMC) and immortalized the primary cells. METHODS: The primary culture of HUASMC was performed using the primary-explant method, and the effects on the cells' growth characteristic by different culture mediums were conducted. Primary cultures of HUASMC derived from normal human umbilical arteries were transfected with recombination plasmid containing human telomerase reverse transcriptase (hTERT) open reading frame. The transfected HUASMC was analyzed for cell biological characteristics, and growth characteristics, and specific alpha-actin. RESULTS: Highly purified primary HUASMC were gained and could be propagated about 10 population doublings (PD). After PD 4, the cells lost the proliferation ability gradually. However, the transfected HUASMC could be propagated at least 20 population doublings and could express specific alpha-actin. CONCLUSION: The hTERT transfected HUASMC could keep the basic biological characteristic of smooth muscle cells and prolonged life span at least 20 PD. It might be a good cell model for the study of cardiovascular disease, myoma of uterus and chronic graft disfunction. It might also bring the hope to engineered vascularization in the future.