两种不同来源成肌细胞存活率的比较研究

A Comparative Study on Viability of Myoblasts Derived from Two Different Tissues in vitro

目的测定和比较两种不同组织来源的骨骼肌成肌细胞的存活率,为组织工程化骨骼肌种子细胞的选择提供参考。方法将两种不同组织来源的第3代成肌细胞接种到24孔板和96孔板中,在1、3、5、7、9d将两种组织来源的成肌细胞分别从24孔板中随机取出5个孔和从96孔板中随机取出10个孔的细胞。将从24孔板取出的细胞置于细胞活力分析仪行细胞存活率的测定。将从96孔板取出的细胞采用MTT法检测细胞增殖活力,测吸光值(OD)。采用细胞存活率的平均值和OD值绘制细胞生长曲线,了解细胞生长情况。结果从骨骼肌中培养的成肌细胞,MTT法测得7d的OD值最高,为0.8153;细胞活力分析仪测定的细胞存活率7d时也最高,为92.06%。采用骨髓间充质干细胞经5-Aza诱导所得到的成肌细胞,MTT法测得7d的OD值最高,为0.6358;细胞活力分析仪测定的细胞存活率7d时也最高,为67.77%。从骨骼肌中培养的成肌细胞的存活率高于骨髓间充质干细胞经5-Aza诱导所得到的成肌细胞存活率(P<0.05)。结论从骨骼肌来源的成肌细胞更适合作为组织工程化骨骼肌的种子细胞。

Objective To compare the two ways so that conclusion is drawn which way is better for supplying seeding cells of skeletal muscle in engineering tissues. Methods The third generation myoblasts were mixed into cell suspension and inoculated in 24 orifices and 96 orifices. Five poroeytes were got randomly from 24 orifices in 1, 3, 5, 7, 9 d respectively, ten poroeytes were got randomly from 96 orifices in the 1, 3, 5, 7, 9 d respectively. Cell viability to 5 poroeytes was determined by the analysator of cell viable. While cell viability of the third generation myoblasts to 10 poreeytes were determined by methyl thiazolyl tetrazolium （MTT）, absorbanee was measured to determine the value of optical density （OD）, finally drawed cell growth curve of myoblasts by using mean value of cell viability and OD so as to comprehend the state of cell growth in vitro. Results To the myoblasts which derived from skeletal muscle, the highest OD value was 0.815 3 by MTT in the seventh day, the highest cell viability was 92.06% by the analysator of cell viable in the seventh day. To the mesenehymal stem cells （MSC） of the second generation were induced by 10 μmol/L 5-Aza, the highest OD value was 0.635 8 by MTT in the seventh day, the highest cell viability was 67.77% by the analysator of cell viable in the seventh day. Conclusion It proved that myoblasts derived from skeletal muscle are more fitable for the source of seeding cells of tissue-engineered skeletal muscle.