Serum-Free Conditions

Introduction

Most of us want to be serum-free or move in this direction, but transitioning to a serum-free media can be a daunting task. Some are struggling with how to plan their transition to serum-free conditions. Some have already transitioned to serum-free but are not getting the results that they want or that they achieved with serum containing media. Still others are stuck in the process of transitioning with cells that just won’t cooperate in serum-free conditions. If this describes your current cell culture situation, you are not alone. Please join us for our Ask the Expert session on cell culture in serum-free conditions.

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Question 1

Can I switch back and forth between Serum Free and serum containing media for short periods of time for experimental reasons?

Hi, This is a great question since sometimes for experimental reasons you might want to do this. Cells that are grown in a serum containing media can be transferred to a serum-free medium, D-PBS (or a serum-containing medium like D-MEM without the serum added) for about 4 hours without hurting them. Remember, if you want … Continued

Question 2

I have a vero cell line that I would like to transition to serum free media, can you give me some advice on where I should start.

This is a good question and those of you what are using VERO cell lines are lucky. GIbco’s VP-SFM is a media designed for growing VERO cells in suspension. VP–SFM is a serum-free media that requires little or no adaptation for VERO cells. The media will also grow other types of cells such as BHK-21 … Continued

Question 3

I have an insect cell line, Sf21 and I need to transition to serum free medium. I have tried to do this before but the growth rate decreased from 24 to 50 hours. I have new cells so what should I do.

Insect cells, Sf21 or Sf9 or HighFive have special growth requirements. Remember the media is not CO2 dependent and the optimum temperature is 27-29C with 95% relative humidity. Insect cells also seem to be special because they have a memory of their conditions and if something is wrong they don’t recover. Also always grow the … Continued

Question 4

I got a growing 293 line grown in suspension in Freestyle-293 from a collaborator. I spun them down, resuspended in fresh Freestyle at 3E5/ml, and incubated at 100rpm 4 days. Cells were at 5E5/ml on day 3 and 4E5/ml day 4. I’m not seeing a lot of death but they don’t seem to be growing. Any suggestions?

I guess the first question is…were the cells growing in the lab of the collaborator and did you switch the growth method. Use a shaker flask without baffles. Also the best thing to do in the future is to split the cells into your media instead of pelleting them. I try not to pellet cells … Continued

Question 6

We are having a hard time culturing primary multiple myeloma CD138+ cells (Isolate from patient bone marrow) and maintaining decent viability in 24-48h. We added 15% patient serum in medium and viability improved a little but some sample still do not have great in viability (30-50% dead in 24h).Would you give me some advice to improve viability?

This is a difficult question to answer without more information. The low viability may de due to the isolation process or/and the culture process. Below is a link to an article in the journal Blood that you may find helpful. http://bloodjournal.hematologylibrary.org/content/103/8/3175.long Total: 0 0 0 0