我也是做缺血缺氧，复氧，急性缺血，开始全部用的原代，那个累啊。
h9c2建议用低糖养，否则高糖刺激对细胞线粒体有影响，具体参考CVR的一篇文章，Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species
还有缺氧我用的ogd液。否则会有干扰，另外h9c2不如原代耐受缺氧，原代的还是比较好，因为事实上心肌就是一直工作中相对低氧缺血的环境中作者: qiangren789 时间: 2013-1-8 10:06
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol: The myoblastic population will become depleted rapidly if the cultures are allowed to become confluent.
To prevent loss of myoblastic cells, cultures should be subcultured before they become confluent, and the line should be recloned periodically with selection for myoblastic cells.
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase作者: 阿司匹林 时间: 2013-1-8 10:06
我也是做缺血缺氧，复氧，急性缺血，开始全部用的原代，那个累啊。
h9c2建议用低糖养，否则高糖刺激对细胞线粒体有影响，具体参考CVR的一篇文章，Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species
还有缺氧我用的ogd液。否则会有干扰，另外h9c2不如原代耐受缺氧，原代的还是比较好，因为事实上心肌就是一直工作中相对低氧缺血的环境中
