Hi everyone,
Recently, I bought a new cell line named Tenocytes from a company.
https://www.angioproteomie.com/ccp6601-human-tendon-cells-28tenocytes29-cap-0041.htm
I followed the manufacturer's instructions and used their medium and coating buffer.
However, I observed that the cell was not attached to the bottom, as shown in the pictures I attached below.
As you can observe here, I saw all cells are still alive. However, they do not attach to the bottom.
I would greatly appreciate your suggestions or any advice for my experiment.
Best regards,
Did you use a culture dish already pre-coated with something? Maybe the coating was ineffective.
How did you determine that "all cells are still alive"? Based on morphology of the cells in the image, one could assume that the cells are on the verge of dying. Did you try a Live/dead assay or staining with annexin/PI or annexin/7-AAD to show viability?
Dear Tina Trinh ,
I would highly recommend to get in tough with the company asap, since they are stating this at the end of the link you have provided:
"Due to the sensitive nature of primary cells and cell lines, all quality related issues about the cells products must be reported back to us within ONE month period after receiving the products, no quality warranty (i.e. replacement of cells) will be provided after the ONE-Month period. Thank you for your understanding."
They should check their batch of the cell stock and give you maybe more clear thawing protocol.
We are usually thawing (different cells) like this:
Thaw the cells 30-60s with in a 37°C water bath (ice should/must still be visible which will keep the cells still cool). Than we are are transferring the cells into 10 ml pre-warmed medium I do use a pipet for that set to 800 µl. After 3-4 pipetting steps adding warm medium into the vial and removing than cells an the medium into the 10 ml reservoir, I spin the cells down (5 min 250 x g) aspirate the medium and plate the cells in new medium into the cell flask or dish. On the next day the medium is exchanged to get ridge of the rest of the DMSO.
Best wishes
Soenke
Dear Sönke Weinert and Alexander Otahal,
Thank you for your suggestions.
I have contacted the company and received their confirmation.
Their cell from the same batch is attached well, and there is no problem.
They also revised my protocol, and there is no difference between their protocol and my procedure.
The only reason may be that the delivery causes cell death.