A Modified Culture Method for Porcine Pulmonary Trunk Smooth Muscle Cells / 华中科技大学学报(医学版)

ABSTRACT

The culture method for porcine pulmonary artery smooth muscle cells (PASMCs) established previously in our laboratory was modified as follows: (1) PASMC mixed medium (PASMCMM) containing 400 ml/L PASMC conditioned medium and 100 ml/L fetal bovine serum (FBS) was substituted for M199 medium containing 100 ml/L FBS; (2) When the PASMC was confluent by 90 %, the subculture was done at the time between the phase of logarithmic growth and stagnation phase of the cells; (3) The method of subculture: The cells were dispersed with 5 to 8 drops of a mixed digestive solution per flash, then M199 medium containing 100 ml/L newborn calf serum was added to terminate the digestion. The cells attached to the substrate of the flashes. On the 2nd day, the media were changed by PASMCMM. The cultured PASMCs by this method were characterized by rapid growth and good dispersion, etc.. The doubling time of the cells (45.6 h) by this method was 11.2 h shorter than that (56.8 h) by the previously established method in our laboratory.The modified culture method for PASMC was more simple, practical and effective, so it can be used as an effecrive method for rapidly harvesting large quantities of PASMCs.