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FAQs

Why is my isolation efficiency very low compared to my current medium?

There are a multitude of potential explanations for such a situation, in variably we need a full description of the procedure you followed. Key questions include what is your current medium? Does it contain FBS or BPE? Was the CELLnTEC medium tested in side by side comparison to your current medium? Did you use trypsin in your isolation protocol? In this case low isolation efficiency could be due to over-trypsinization, previously the trypsin reaction was inhibited by proteins from FBS or BPE. Other reasons for over- trypsinization include high trypsin concentrations or a too long trypsin treatment. Refer to the specific FAQ above where we explain in detail the different options of detaching cells. For more detailed info, email scientist@cellntec.com

My previously isolated cells, growing in my current medium did not grow very well when switched to the CELLnTEC medium. What could be the problem?

Primary cells can quickly adapt to specific cell culture media. Hence when switching a cell culture to a different medium, there is a need to wean your cells off the old medium. This is accomplished with a step-wise serial dilution of the old medium with CELLnTEC medium over a period of one to four weeks, depending on the severity of the transition and the proliferation rate of the cells. See detailed protocols listed above.

I am developing therapies based on adult stem cells. Is the CELLnTEC medium produced as a therapeutic grade product?

Basal media are manufactured in an ISO 9000 certified facility according to cGMP guidelines. However these media are only intended for research applications. When medium with greater certification is required (for example in phase I clinical trials), custom formulations with more highly certified components and expanded quality control procedures can be manufactured. In all cases, CELLnTEC must begin by working with the customer to determine the specifications needed for their application.

Why didn’t my cells grow after passaging them?

A very common problem researcher’s encounter when switching to a serum free or low serum medium is that the trypsin is not deactivated with the addition of medium. Over digestion with trypsin can irreversibly damage the cells, thus detachment must be closely monitored under the microscope. CELLnTEC recommends using a milder enzyme such as Accutase for the passaging of cells. A protocol is found on our resources section.

I have early passage cells growing, can I swap them directly into the CELLnTEC medium?

No, cells grown in one medium are adapted to this medium and therefore it is necessary to be weaned away from the old medium to the new medium. This is best done in a step-wise decreasing dilution of the old medium over a period of one to four weeks, depending on the severity of the transition and the proliferation rate of the cells. For a four week approach this can be done with the following dilutions (new medium / old medium): 20 / 80%, 40 / 60%, 50 / 50%, 60 / 40%, 80 / 20% and 100 % new medium. For a two week approach the following dilutions can be used (new medium / old medium): 25 / 75%, 50 / 50 %, 75 / 25% and 100% new medium. For a one week approach the following dilutions can be used (new medium / old medium): 25 / 75%, 50 / 50 %, 75 / 25% and 100% new medium, in this case the change is made much quicker than in the two week approach and additional you should use a higher seeding density (10 to 20% more than normal) to be sure that enough cells will attach and survive to have the best density conditions for the further culture. Please keep in mind that changing medium sometimes can lead to changes in cell morphology or growth behavior of the cells.