Some more details would be beneficial. Is it a retrovirus producer cell line? What is the plasmid encoding for - envelope protein or something else - capsid etc? I have worked with a retrovirus HEK293 producer cell line before and can give you some details.

 You can please follow the links below to obtain certain crucial informations:-

1. http://www.ncbi.nlm.nih.gov/books/NBK19423/

2. http://www.sciencedirect.com/science/article/pii/S095816690000080X

3. http://www.invivogen.com/docs/Insight_201004.pdf

4. http://web.stanford.edu/dept/EHS/prod/researchlab/bio/docs/Working_with_Viral_Vectors.pdf

5. https://research.cchmc.org/translationalcores/vector-production/gmp-vector-production

Dear Sandip Chakraborty this articals very useful for my research ant thank you very much for your interest.

Dear Rahul Keswani, thank you for answer to my question. Actulally, I want to use clonetech Lenti-X HTX Packaging System and their specific plasmids (pLVX-Puro Vector). If possible, I try to create successfully packaging HEK293 cell clones and freeze them until my next work. I want to learn, when I thaw and growth appropriate condition, does these cells, produce effective viral particles.

Hi Alper,

My graduate lab used the Retro-X and Lenti-X system pretty commonly. I have used only Retro though. I can suggest a few things right now.

The kit should come with a pLVX with luciferase as well. Use that to make sure that your producer cell line can efficiently produce viral particles. You can use their Lenti viral quantification kit as well as perform some transduction experiments and measure light units.

As far as the actual cell line goes, the first transfection is important and make sure you follow all their steps to the letter. These cells can be frozen and thawed multiple times but keep enough stocks frozen. I never passaged my producer cell line more than 25 times. The kit suggests not more than 10, I think. 

We used regular growth media - DMEM (high glucose, pyruvate) + 10% FBS with no antibiotic. We noticed that the type of serum we used did change viral performance drastically. So once we got the cells growing, we stuck to the same serum lot for that particular cell line (not just the brand, but the same lot#).

We got a good number of viral particles upon 3-4 days of passage - ~1-2 x 10^10 viral particles/ml. Do not replace media when obtaining viruses. Keep it in the same media for 3-4 days. 

Use the envelope plasmid when you want to test for performance. I also suggest my 3 papers wherein I made hybrid vectors using non-enveloped viral particles with polymers and lipids. you can use those protocols to test for performance as well and it will be cheaper as well.

http://pubs.acs.org/doi/abs/10.1021/mp300561y

http://www.sciencedirect.com/science/article/pii/S0168365914004696

http://www.sciencedirect.com/science/article/pii/S0168365915002333

If you have any specific questions, let me know.

 Hi Rahul, Thank you for your suggestions and valuable data. They are very helpful for me.