In the last 20 years, thanks to the adaptation in suspension of several cell lines and to the development of more efficient transfection agents (eg lipofectamines) and culture media, the scientist were able to obtain amazing progresses in the performance of mammalian expression systems,
As I already reported in the ProteoCool n° 29 and in the tips of the week#8 , modern cell lines, as ExpiCHO
and Expi293, are able to produce high levels (up to 300-500mg/l) of recombinant
antigens and/or antibodies in the culture surnatants also in shaking flask culture.
The availability of an orbital shaker able to maintain 5-8% of CO2
atmosphere is essential during cell propagation and maintenance.
Differently to bacteria, the cell lines do not require high shaking
rates (generally 120-125rpm) but are more prone to contamination, therefore since
the cleanliness is essential, the use of sticky pads, those can be easily
removed and sainted with ethanol or isopropanol solutions is preferable respect
than the conventional flask clamps.
To my knowledge Eppendorf S41i (previously New Brunswick) and the Infors multitron are the mostly used for mammalian cell cultures.
In this presentation I would like to share with you, my experience with the Eppendorf S41i
In our lab we have 2 Eppendorf S41i shakers:
- 1st setted at 37°C- 8% CO2 and 120rpm for propagation of both Expi293 and ExpiCHO and for transfection of Expi293 cells;
- 2nd setted at 32°C- 5%CO2 and 120rpm for the transfection
of the ExpiCHO cells;
The first shaker was acquired in 2016-2017 while the second in the 2020.
Our older S41i shaker carry a platforms characterized by a smooth surface and the sticky pads attach well directly to its surface
while our newest S41i shaker carry a platforms characterized by a rough surface and the sticky pads don’t stick well to its surface.
Eppendorf sell a Stickpad ADAPTOR KIT which contains 2 metal smooth plates
but this kit seems to be designed for a different shaker format since
the sizes of those plates are not appropriate to cover the entire area of the shaking platform.
You can use the fixing screws supplied with the conventional flask clamps or wtih the Stickpad ADAPTOR KIT to drilling and fixing the sticky pads directly on the black shaker platform.
Standard Washers with 6mm in diamteter
those can acquired in the most hardware stores /e,g Leroy Merlin, Obi, Bricoman, Brico ) to ensure the sticky pad integrity and extend the life time of this solution.
N.B I'm happy to share with you that, as responce to this post, Eppendorf specify that all the platforms actually produced are chatacterized by a smooth surface and the sticky pads are able to attach directly to it.
Hello from Hamburg :) Very nice and informative blog! We are happy to hear that you are using the Eppendorf S41i and – beside the sticky pad issue – seem to like it! I would like to comment on the platform issue with sticky pad. A growing amount of our suspension cell culture customers use the sticky pad. Because of this, in 2020 we improved the platform surface to make it better compatible with sticky pad. The new S41i universal platform consists of hard coated, anodized black aluminum, a suitable surface for direct use of sticky pad and still resistance to high temperature disinfection. So, the new one has a smoother surface. The prior one with a rougher surface can still be used, there is no quality issue, but when using sticky pads, the adapter kit is required for best hold, you can use e.g. test tube rack or clamps next to it to not loose space. So, whenever you buy a new one, you get the smooth one which is compatible with sticky pad! So, it is just vice versa. Best, Ines from Eppendorf
ReplyDeleteDear Ines
Deletethanks a lot for the feedback
it is important for all the present and future user to know that now the universal platform of S41i shakers has a surface compatible with sticky pads since sticky pads are essential for the culture of mammalian cells in suspension.
thanks again
Manuele