一种改良的猪肺动脉干平滑肌细胞培养方法

A Modified Culture Method for Porcine Pulmonary Trunk Smooth Muscle Cells

在本室已建立的肺动脉平滑肌细胞 (PASMC)培养方法的基础上作了改进 :1用含 40 0 ml/ L PASMC条件培养液和 10 0 ml/ L胎牛血清 (FBS)的 PASMC混合培养液 (PASMCMM)代替含 10 0 m l/ L FBS的 M199培养液 ;2传代时间选在 90 %长满 ,即 PASMC处在对数生长期末而停滞期前 ;3用 5～ 8滴 /瓶混合消化液传代 ,然后用含 10 0m l/ L小牛血清的 M199培养液终止反应 ,并使其贴壁 ,第 2 d改换成 PASMCMM。经此法培养的 PASMC具有生长快、分散更均匀的特点。细胞倍增时间 (4 5 .6 h)较改进前 (5 6 .8h)缩短 11.2 h。改进后的培养方法更加简便、实用、高效。因此是一种快速、并可大量获取优质

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 effective method for rapidly harvesting large quantities of PASMCs.