Rabbit periodontal ligament stem cells

1) Prepare the necessary materials for cell culture in advance according to the instructions for cultivation conditions before receiving the goods.
2) After receiving the cells, please check the condition of the shipped culture bottles. If any damage, liquid overflow, or cell contamination is found in the culture bottles, please take photos and contact us promptly.
3) After receiving the goods without any appearance damage, disinfect the T25 bottle with 75% alcohol and place it in a 37 ℃ incubator for about 2-3 hours to wait for the cells to stabilize.
4) After stabilizing the cells, confirm the cell growth status under a microscope, determine the cell density, and take clear photos at 200x and 400x magnification. At least 2 photos should be taken to record and provide feedback on the cell status as evidence to prevent after-sales issues. If there are no photos or feedback, it is assumed that the cell quality is acceptable.
5) Adherent cells: During transportation, adherent cells may detach due to bumps. If detachment or clumping growth is found, T25 bottles can be placed in a 37 ℃ incubator for about 2-3 hours. Then, the culture medium and non adherent cells in the bottle can be extracted and centrifuged at 1000rpm for 5 minutes. Discard the supernatant and resuspend it, then inoculate it into a new culture bottle or dish. Add the newly prepared complete culture medium according to the instructions for cell culture conditions.
6) Suspension cells: Place the T25 bottle in a 37 ℃ incubator for about 2-3 hours, then extract the culture medium and cells from the bottle and centrifuge at 1000rpm for 5 minutes. Discard the supernatant and resuspend, then inoculate into a new culture bottle (add complete culture medium prepared according to the instructions for cell culture conditions).

Notes:
A. Passage ratio:
One T25 bottle is transferred to two 6cm culture dishes, or two T25 bottles are equivalent to a ratio of 1:2
1 T25 bottle transferred to 1 10cm culture dish or 1 T75 bottle is equivalent to 1:3
B. Due to the limited number of passages and activity of primary cells, it is necessary to plan the time for culturing and conducting experiments after receiving the cells, in order to prevent the experiment from starting only when the lifespan of the primary cells has reached its maximum limit. At this time, the primary cells are likely to have poor growth ability, morphology, and status, so it is best to make predictions and judgments in advance.

It is recommended to ship primary cells at room temperature for a more stable delivery status of live cells. The T25 bottle packaging of primary cells we ship usually has a cell density of around 80% and is filled with conventional culture medium. Customers can decide whether to collect and continue using it for 3-4 days or to remove and discard it according to their own experimental plan.