MC3T3-E1 mouse embryonic osteoblast precursor cells
Components
Operation steps for cell culture
1. Aspirate the medium for use, add 3-5 mL of PBS, and gently shake the flask to rinse the cells;
2. After PBS is cleaned, add 1 mL of 0.25% trypsin-0.53 mM EDTA, gently shake the dish to allow the trypsin to soak the cell surface, and incubate the flask in a 37-degree incubator. (The cells are sensitive to digestion, and excessive digestion may seriously affect The state of the cell causes the cell to pass on death or float. After the cell is digested, the cell gap becomes large but does not fall off. It can be terminated when it is gently blown down. It is forbidden to digest until the cell is completely floating. Mix the cells as gently as possible. Blowing out a lot of bubbles.)
3, add 6-8ml complete medium to terminate digestion, gently blow the cells to mix;
4. Centrifuge the mixed cells at 1000 rpm (about 150 g) for 3 min, discard the supernatant, and resuspend in a 37-degree incubator with fresh medium to continue the culture;
Passage ratio: Different cell growth rates vary, and the specific passage ratio depends on the cell growth rate, and most cells are suitable.
1:3-1:4 Passage, slower growing cells can be passaged 1:2.
Cell cryopreservation
1. Cryopreservation solution: 92% complete medium + 8% DMSO (can be selected according to laboratory conditions)
2, cooling step: 4 degrees 10min, -20 degrees 2h, -80 overnight after storage of liquid nitrogen.
| Catalogue No. | C1047 |
| Product Format | a T25 flask |
| Culture Properties | Sticking |
| Complete Growth Medium | 89% H-DMEM + 10% FBS + 1% double antibody |
| Atmosphere | Air, 95%; carbon dioxide (CO2), 5% |
| Application | Cells and cancer research |
| NOTE | FOR RESEARCH USE ONLY. |
Components
| Item | Specifications |
| a T25 flask | 2X10 6 |
| Manual | 1 copy |
Operation steps for cell culture
1. Aspirate the medium for use, add 3-5 mL of PBS, and gently shake the flask to rinse the cells;
2. After PBS is cleaned, add 1 mL of 0.25% trypsin-0.53 mM EDTA, gently shake the dish to allow the trypsin to soak the cell surface, and incubate the flask in a 37-degree incubator. (The cells are sensitive to digestion, and excessive digestion may seriously affect The state of the cell causes the cell to pass on death or float. After the cell is digested, the cell gap becomes large but does not fall off. It can be terminated when it is gently blown down. It is forbidden to digest until the cell is completely floating. Mix the cells as gently as possible. Blowing out a lot of bubbles.)
3, add 6-8ml complete medium to terminate digestion, gently blow the cells to mix;
4. Centrifuge the mixed cells at 1000 rpm (about 150 g) for 3 min, discard the supernatant, and resuspend in a 37-degree incubator with fresh medium to continue the culture;
Passage ratio: Different cell growth rates vary, and the specific passage ratio depends on the cell growth rate, and most cells are suitable.
1:3-1:4 Passage, slower growing cells can be passaged 1:2.
Cell cryopreservation
1. Cryopreservation solution: 92% complete medium + 8% DMSO (can be selected according to laboratory conditions)
2, cooling step: 4 degrees 10min, -20 degrees 2h, -80 overnight after storage of liquid nitrogen.
Tips :
1. After the cells are transported, some cells are broken due to temperature changes and violent collisions, which is a normal phenomenon. The adherent cells can be digested, and the suspended cells are directly mixed and collected, centrifuged at 900-1000 rpm (about 150 g) for 3 min, and the supernatant is discarded. The cells were resuspended in 5 ml of PBS, centrifuged at 900-1000 rpm (about 150 g) for 3 min, and resuspended in fresh complete medium to inoculate the new flask. The second PBS resuspension is to remove the debris. If there are few fragments in normal times, the step of resuspending the PBS can be omitted during the passage; if there are many fragments, it is recommended to wash the PBS several times.
2. When the cells grow unevenly, the cells can be digested and dispersed, and then added to a new medium for re-inoculation or passage.
3. When the cells grow slowly, you can choose to increase the serum concentration culture (up to 20%), or you can select the passage cells to continue the culture according to the cell growth state.
4, the difference in adherence of different cells is relatively large, so the difference in digestion time is large, 20s-10min is possible, specifically the cells are digested to separate from each other but not fall off, and can be gently blown down, it is strictly prohibited to digest the cells completely float. Customer digestion caused by excessive cell death, floating, slow growth, does not provide free service.
5, dry ice delivery are two, the customer first recover one, if the recovery fails, contact us in time and recover the second branch under our guidance. If two customers at the same time of recovery are poor state, we do not provide free service.
6. When the cell state is normal, the cells should be cryopreserved as soon as possible, and a frozen cryopreservation effect should be randomly selected after cryopreservation. I SHALL NOT responsible for the customer frozen cell death, cell death frozen client does not provide free service.
Notice:
If the customer receives any questions from the cell, please call us in time. If there is no phone call within 1 week after receiving the cell, or other forms of return visit, the default is no problem with the cell quality. After any problems, no free after-sales service will be given. The cells are only provided once after free sale. If they are re-cultured after re-issue, the death will not be reissued free of charge, except when the cells are dead or insufficient in density.
Gun Safe,Gun Safe Box ,Mechanical Gun Safe Box,Gun Safe Box With Combination Lock
Ningbo Zhaomu Electronic Commerce Co., Ltd. , https://www.bofonhome.com