I mentioned to to Chris already, but we use nothing but HyClone SFX-Insect powder. We make 20-30 L batches, sterilize with a large peristaltic pump and a disposable Millipak filter from Millipore. We never have contamination problems that are due to preparing our own media from powder. We buy 10L bottles of powder. This media works great for baculovirus, stable cells, Sf9, Sf21, Tn5. We prefer it over Invitrogen (why have separate media for Sf9 and Tn5 cells?), and it's cheaper too! We no longer use serum media for anything (virus production and amplification, all goes fine in serum free. Serum helps storing viral stocks for long periods). It's ~$26 per L, plus $1-$2 in filter costs (a $30 filter will sterilize ~30 L of media). So it comes to about ~$28/L, I haven't priced out alternative products recently, but I think liquid from Invitrogen runs around $50/L?
We also use the peristaltic pump for tangential flow filtration, it's pretty heavy duty (tubing is like 5/8" thick). I think that help make the filtering efficient. Bottle top filters won't work because they foul to quickly, same is try of 4.5 cm filter discs that you can insert into an in-line filter, they clog after 1 L or so. You need the 'asymmetric' filter material offered by Millipore in the Millipak or Steripak filter (the later is the filter you see attached to a MilliQ water dispenser, it can be autoclaved 3X they claim) Hope that helps, Nat On Mon, Jun 7, 2010 at 10:59 AM, Tommi Kajander <tommi.kajan...@helsinki.fi> wrote: > Hi, well, actually this is availalbe as powder (HYClone), as it says on the > page.. (we have it made in our media kitchen on a regular basis, but > not for huge scale... so i dont know about the prices..) > > https://www.thermoscientific.com/wps/portal/ts/products/detail?navigationId=LA11074__10347&categoryId=82051&productId=11960716 > > HTH, > Tommi > > > On Jun 7, 2010, at 5:52 PM, Dima Klenchin wrote: > >>> We have happily made a transition last year from using Invitrogen's >>> SFM medium and cellfectin to Insect-Xpress (Lonza) and >>> polyethyleneimine for transfection. We are moving several protein >>> targets to large-scale cultures and would consider cost-cutting >>> alternatives. For example, Invitrogen and Thermo Sci both offer media >>> in powder form at an attractive price. Has anyone made a systematic >>> comparison of these media? Any particular recommendations regarding >>> powder media? >> >> Last I checked, no manufacturer offered serum-free powedered medium. >> Fetal calf serum is very expensive - once you count in its cost, the >> overall price of the media becomes comparable. >> >> If your protein is secreted, serum-containing media are not even a >> realistic choice. If it is intracellular, buying dry medium and >> adding FBS work well: is a lot more hassle, is about 20% cheaper, >> and usually gives slightly higher titers and expression levels. Of >> all the common media we compared, TNM-FH works best for us with Sf9 >> cells. Switching to dry medium also means some initial investment: >> bottles, filtration equipment and filters. So it all pays off only >> of you need large scale cultures long-term. >> >> If you are aware of a dry serum-free insect cells medium, please >> let me know - I'd love to try it. >> >> Dima >> > > Tommi Kajander, Ph.D. > Structural Biology and Biophysics > Institute of Biotechnology > University of Helsinki > Viikinkaari 1 > (P.O. Box 65) > 00014 Helsinki > Finland > p. +358-9-19158903 > tommi.kajan...@helsinki.fi >
