TS Answer: the MTOX1010 cells are intended for use as both control cells for HepaRG™ knockout cells as well as for a wide variety of liver cell based assays. As such we do not have a passage number limit but a minimum culturing period. Cells should be differentiated for a minimum of 14 days, exchanging medium twice weekly, before our suggested assays (see protocol attached).
Please note that the MTOX1010 HepaRG™ 5F Control Cells as well as the MTOX1011 PXR Knockout HepaRG™ Cells are patented, their use is strictly limited, and you are not allowed to passage them further. If you wish to do so, you would need to purchase a license.
HepaRG™ 5F and Knockout Limited Use License
These cells are derived from HepaRG® cells and are protected by patents; by opening this package, you agree to abide the terms of this Limited Use License as follows. You will consider the cells as a disposable product to be destroyed upon conclusion of a study or experiment; will use them only for in-vitro experiments and only at the facility of their receipt; will not propagate, amplify, reproduce, clone, or make any other use of them after a study is concluded; will not produce or manufacture commercial products or cDNA libraries or cell stock from the cells; will not use them for any study of a duration exceeding 20 days; and you will not license, resell or transfer them to anyone outside your organization for any reason. If you are unwilling to accept the terms of this LIMITED USE LICENSE, do not use the cells and return them to Sigma-aldrich for credit. Violators of this LIMITED USE LICENSE will be prosecuted to the fullest extent of the law.
You can find additional information via the following link:
https://www.sigmaaldrich.com/life-science/adme-tox-assays/license-agreements.html
The Technical Bulletin link below describes the confluency determination and growth rate:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/530/047/mtox1010bul.pdf