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Next Subject 		Spinner flask CHO cell suspension culturemanishkibaat2009-02-11Click here to register.
I am trying to grow the CHO suspension cell culture in a wheaton 500 ml spinner flask. i am using JRH-SAFC CD CHO medium with 1x10E5 cell/ml seeding density. Initially i am starting with 200 ml culture for four days & then adding 200 ml more fresh medium. RPM is throughout 100. The problem is that cells are not dividing fast & i am not getting the expected count. Count after 7-8 days remains 6-7 x 10E5 cell/ml. Some times cell viability also dips after 3-4 days. Measured culture pH always remains between 7.0-7.2.
I have also tried different volumes, different seeding densities & CO2 percentage.
It will be highly acknowledged if somebody suggest me the solutions orn else some suggestions.

Thanks & regards
Manish Acharya
 
Re: Spinner flask CHO cell suspension culture
Jryan2009-03-01Click here to register.
Are the cells and medium exposed to fluorescent light either for long periods during storage or while in the spinner? If so, you may be seeing toxicity from hydrogen peroxide produced by the light interacting with medium components.
 
Re: Spinner flask CHO cell suspension culture
manishkibaat2009-03-02Click here to register.
Firstly thanks for the interest. but further ti teel that its not the case of long light exposure. Cells remain in dark incubator while in spinner flask & medium also remains at 2-8 degree in refigerator when not in use.Please suggest some other solution. Thanks
 
Re: Spinner flask CHO cell suspension culture
BAJ2009-03-16Click here to register.
I find CHO cells do better when split to 1.5e5c/ml or 2e5c/ml. CHO cells should be hardy and have a doubling time of 24 hours or less. Have you grown them in other vessels and do you see better growth there? Have you grwon them in other media? It is possible the basal medium is not appropriate for this particular cell line; are other media an option? Both pH and rpm should be fine for CHO cells.

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