Question Topic:

Serum-Free Conditions

Sponsored by: Life Technologies
Session ends: Closed
Answers by: Timothy Fawcett, Ph.D. Director of the BioTechnical Institute of Maryland (BTI)

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.
Ask the Expert Image

This session is sponsored by
Life Technologies

 


Questions & Answers

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» Read More

Can you recommend good and inexpensive Serum-Free Insect Cells media? I know that words good and inexpensive can not often be used in one sentence, but one can always hope… I am currently growing SF9 cells in SFM from Allele Biotech and having good results in maintaining cell culture and protein expression. However it is quite expensive($70 for 1L). Can anybody say anything about BD Bioscience BaculoGold or Corning Mediatech SF media? Any other media suggestions would be very much welcome and appreciated.

Serum-Free media are expensive but do outperform serum containing media. To be honest with you I have used Gibco’s Sf900. Sf900II and Sf900III and several other manufactures serum-free media and the new Sf900 III media is the best I have used. The problem you will face if you switch is adapting your cells to a […]» Read More

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 […]» Read More

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 […]» Read More