BHK 21 cell lines culture

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harish.kuppam
harish.kuppam's picture
BHK 21 cell lines culture

we are culturing BHK 21 cell lines for the production of FMD vaccine. we find difficulty to come across the massive cell death in bulk culturing of cell lines using fermentors though we are using all the aseptic conditions and filterations. can some one please suggest me how to get rid of this problem. i need an immediate help from u guys.

Shubhangi
Shubhangi's picture
Hi Harish

Hi Harish
There is no direct answer to your question,only suggestions......
You have mentioned that you are using aseptic conditions, still there is always chance of microbial contamination. This could be easily found by microbial testing in QC or by microscopic observation.
Manytimes in fermentors the problem could be improper washing- chemical contaminant, or fluction in temp,pH, O2/CO2 etc. Also check your media, is it a new lot?

 

harish.kuppam
harish.kuppam's picture
we have made 15 lots already

we have made 15 lots already and this problem araised all of a sudden from the 16th lot and some times the cells are becoming non-viable. for checking sterility we are using SCDM, FTM and NA. we use Minimum essential hi media with 8% ABS along with streptomycin, kanamycin and bezyl penicillin. till some passage levels it comes nicely and when we go for higher production we are getting either clumps or cell death. clumps look like a group of cell gathering together and forms a shell around them. we are using 0.2 micron filters and highly steriled fermentors.

asif123
asif123's picture
Dear  harish.kuppam  

Dear  harish.kuppam  
This is my first experience to work with BHK-21 cell line at bioreactor level (1L) I don’t know about the saturation (saturation by air or O2), DO setting for growth and production and how to manage the perfusion rates. At present I am using McCOYS 5A media but I don’t know will it support at the bioreactor level or not.
I will be very grateful of your help.
 
Thanks in advance
 

Qaiser
Qaiser's picture
Hi Harish ...howzzz??/have

Hi Harish ...howzzz??/have you got solution or overcome your problem????
Actually M also doing work on BHK cells for mass production ov FMD vaccine I want your concern and Mr.Shubhangi would you like to please Help me to do so and guide me about it????I 'll be thankful to both of you...

girish
girish's picture
saturation with air is

saturation with air is accountable. bcaz if u saturate with air , the partial pressure of oxygen is 21% and it will become 35% in water. so 50% of this saturation point is sufficient for the growth of the cell. If u saturate with oxygen, the partial pressure of oxygen becomes 100% and it can accountable upto 24mg of oxygen concentration which is highly toxic to the cells.

TECHNO WORK
TECHNO WORK's picture
 Have you checked for

 Have you checked for mycoplasma, Which type of FBS you are using.

TECHNO WORK
TECHNO WORK's picture
 Which type of ABS you are

 Which type of ABS you are using?