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Can mycoplasma grow in the absence of cells?

There is evidence to suggest that mycoplasma can survive in the absence of cells, some may even proliferate, but as a rule they are much more viable if cells are present. 

How can I improve Human Hepatocyte viability post Nucleofection on the 96-well Shuttle device?

In addition to proper cell handling and using culture media optimized for long term culturing of hepatocytes (Lonza Basal medium supplemented with BulletKit), we recommend separating the healthy hepatocytes from the dead cells and debris in the...

Do you have data about the in vivo implantation of hepatocytes transfected by Nucleofection? For how long could protein expression be detected?

There is a publication reporting on the successful reinjection of modified hepatocytes into mice.Chen et al. were able to monitor the expression over several days. This group worked with a preliminary test solution for mouse hepatocyte as the...

Is it possible to store reconstituted MycoAlert®/MycoAlert® PLUS Reagent and Substrate?

For optimal assay conditions, reconstituted reagent and substrate should be used fresh within 2 hours after reconstitution.During this time they can be kept at room temperature.For the MycoAlert® Kit: Unused, reconstituted components can be aliquoted...

Are rat and mouse hepatocytes transfected by Nucleofection® still functional?

Yes, transfected mouse hepatocytes show albumin synthesis rates comparable to those of untransfected cells. Moreover the morphology and polarization of rat hepatocytes is not affected by Nucleofection®.

How much sample do you need for accurate results with MycoAlert® Assay? Do you need cells or just supernatant?

The standard protocol calls for taking 2 ml of culture media, spinning down any cells and removing 100 µl of supernatant as sample.Smaller initial aliquots of media may be taken to start with if necessary.

What is the best way to centrifuge Human Hepatocytes prior to Nucleofection?

In order to avoid cell compaction in the pellet and difficulties in cell resuspension, we recommend using round bottom 2 ml vials or 50 ml BD Falcon™ tubes for all centrifugation steps prior to Nucleofection™.

Is it possible to quantify the mycoplasma with the MycoAlert® Mycoplasma Detection Assay?

No, the MycoAlert® Mycoplasma Detection Assay (as well as the MycoAlert® PLUS Mycoplasma Detection Assay) is not quantitative; it has been designed to give a simple Yes or No answer, pertaining to the presence of Mycoplasma.

When can I start induction studies on Human Hepatocytes transfected with the Nucleofector®?

We recommend allowing the transfected hepatocytes to recover in Plating medium for 72 hours post-Nucleofection®.For induction studies (e.g. CYP3A4), the Plating medium is replaced with maintenance medium (William's E without FCS and EGF) and...

When performing the MycoAlert® Mycoplasma Detection Assay, why is it necessary to spin out the cells?

It is highly recommended that the samples are spun down prior to testing. If cells are present in the assay they will contribute to the initial reading producing a higher background. This in turn can decrease the difference between the background and...
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