Jiang Xue-you, Zhang Li, Yu Chen, Jiang Hao, Li Juan
Sichuan Da Xue Xue Bao Yi Xue Ban. 2015 Mar;46(2):301-4.
To seek a simple and effective approach through probing method about primary culture and purification of neonatal rat cardiac myocytes.
Twenty neonatal rats were randomly divided into 2 groups. Group A: the cardiac myocytes were gained by means of enzymic digestion and tubularis-blowing; Group B: the cardiac myocytes were gained by means of enzymic digestion and rotator-stirring. The cardiac myocytes in both groups were purified by means of differential attachment technique and BrdU inhibition and were identified with immunofluorescence staining. Viability was assessed with trypan blue staining and pulsation was assessed under microscope.
The rates of cardiac myocyte's viability, pulsation and purity in group A were (89.90 +/- 2.92)%, (91.30 +/- 2.00)% and (94.90 +/- 1.79)% respectively; The corresponding rates in group B were (94.70 +/- 2.31)%, (95.00 +/- 1.24)% and (95.60 +/- 1.43)% respectively. The rates of cardiac myocyte's viability, pulsation in group B were significantly higher than that of group A, which was statistically different (P>0.05). The rate of cardiac myocytes purity in group B was a bit higher than group A, but which wasn't statistically different (P<0.05). Fibroblasts cultured with BrdU grew weaker and less than those without BrdU.
The method of rotator-stirring and enzymic digestion was better than that of tubularis-blowing and enzymic digestion, which helped to gain cardiac myocyte effectively. BrdU helped to purify cardiac myocytes.
通过探索性方法寻找一种简单有效的新生大鼠心肌细胞原代培养及纯化方法。
将20只新生大鼠随机分为2组。A组：采用酶消化法和吹管法获取心肌细胞；B组：采用酶消化法和旋转搅拌法获取心肌细胞。两组心肌细胞均采用差速贴壁技术和BrdU抑制法进行纯化，并用免疫荧光染色进行鉴定。用台盼蓝染色评估细胞活力，在显微镜下评估搏动情况。
A组心肌细胞活力、搏动率和纯度分别为(89.90±2.92)%、(91.30±2.00)%和(94.90±1.79)%；B组相应的比率分别为(94.70±2.31)%、(95.00±1.24)%和(95.60±1.43)%。B组心肌细胞活力、搏动率均显著高于A组，差异有统计学意义(P>0.05)。B组心肌细胞纯度略高于A组，但差异无统计学意义(P<0.05)。用BrdU培养的成纤维细胞生长比未用BrdU的更弱且更少。
旋转搅拌酶消化法优于吹管酶消化法，有助于有效获取心肌细胞。BrdU有助于纯化心肌细胞。
