PMC-OBC02-COS-Osteoblast Culture kit V-1（成骨细胞培养试剂盒）-骨研究-wako富士胶片和光

发表于由

破骨细胞的骨吸收作用和成骨细胞的骨生成形成了骨代谢平衡。成骨细胞培养试剂盒（大鼠）（PMC-OBC02-COS）含有从大鼠颅骨中分离的冷冻成骨细胞和培养基，可用于成骨细胞和骨生成研究。

破骨细胞的骨吸收作用和成骨细胞的骨生成形成了骨代谢平衡。成骨细胞培养试剂盒（大鼠）（PMC-OBC02-COS）含有从大鼠颅骨中分离的冷冻成骨细胞和培养基，可用于成骨细胞和骨生成研究。
[Protocols]
1-1. Cultured with the 25 cm2 flask
1. Thaw the culture media in a 37°C water bath withgentleshaking.
2. Quickly place osteoblast vial in a 37°C water bath until the contents are thawed.
3. Transfer thawed cells into a 15 ml centrifuge tube containing 10 ml of culture medium and centrifuge for 5 minutes at 4°C at 600 x g for 5 minutes.
4. Remove the supernatant, re-suspend cells in 10 ml of culture medium and centrifuge at 4°C at 600 x g for 5 minutes.
5. Remove the supernatant, and re-suspend the cell pellet in approximay 5 ml of culture medium.
6. Transfer the cell suspensionto 25 cm2 flask and incubate the flask at 37oC under 5% CO2 and 100% humidity.
7. The next day, change the medium.
* Approximay 2-3 days of culture, cells become confluent. For subculture, please refer to the protocol below. Subcultureof the cells can be performed up to passage 2.

1-2. Subculture
1. Subculture the cells when they are confluent.
2. Prepare sterile washing buffer (Hank’s BSS or PBS(-)), and trypsin/EDTA solution. Warm washing buffer in a 37°C water bath priortouse.
3. Rinse the cells with 5 ml of washing buffer twice.
4. Remove washing buffer and then add 3 ml of trypsin/EDTA solution into flask (25 cm2 flask).
5. Gently rock the flask to make sure that the cells are covered by trypsin/EDTA solutionand thenimmediay remove trypsin/EDTA solution.
6. Incubate the flask in a 37°C incubator until cells are compley rounded up (monitored with inverted microscope). Approximay it takes 2 to 3 minutes.
7. Add culture medium to the flask and transfer detached cells to centrifuge tube, and then centrifuge the centrifuge tube at 4°C at 600 x g for 5 minutes.
8. After removing the supernatant, re-suspend cells in culture medium and centrifuge for 5 minutes at 4°C at 600 x g for 5minutes.
9.Remove the supernatant, and re-suspend cells in culture medium. Count cells and plate cells in a new plate or flask (Adjust cell density to the desired experiment).
* Approximay 2-3 days of culture, cells become confluent when seeding density is 30,000 cells / cm2