cell harvesting; use of antibiotics? - (Dec/13/2007 )

Hello everybody,

I have kind of a basic question: I will dissect tendons from animals (mice/rats) and would like to harvest tendon cells thereafter. After cutting out, the tendons are stored in PBS (short termed, approx. 20-30 minutes), followed by enzymatical digestion. Would you add antibiotics (pen/strep) to the pbs to avoid contamination of the culture? work is aspeptic, but i am still worried. would I rather harm the cells by adding pen/strep?

Thanks so much in advance!

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It's not so easy to get sterile cells even if you work aseptic.
I would use peni/strep, unless if you are working in the bacteria/host interaction field.

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Hi Missele,

thanks... this helps a lot. No, I am not working in the bacteria/host field so I will add pen/strep.

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If you do the dissection in a hood and spray the animal with ethanol before dissection, these things can also reduce incidence of contamination.

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Good answer scolix,

It is easy to get clean cells from a "sterile animal" like a rat/mouse without any antibiotics. It can also be achieved from a filthy abattoir when collecting pig/sheep tissue for growing primary SMC's/Endothelial cells.

It a matter of good aseptic technique and TRAINING.

Rhombus

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It is easy to get clean cells from a "sterile animal" like a rat/mouse without any antibiotics. It can also be achieved from a filthy abattoir when collecting pig/sheep tissue for growing primary SMC's/Endothelial cells.

It a matter of good aseptic technique and TRAINING.

Rhombus
[/quote]
Yeah. Pre-cleaning, a lot of washes with saline plus the use of gentamycin usually does the trick for primary cell harvest.

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Thanks everybody... this is of great help. in case someone will read it:
the animals are killed with an overdose CO2 - some claim it will affect cells, some say it is ok.
Do you have any negative experience with CO2 overdose and the primary cell culture?

Thanks!

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When we make mouse embryonic fibroblasts we never use antibiotics and very rarely have a contamination issue (once in the 2 years I've been here). As long as you are careful, you should be fine

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We have not seen any negative effects on primary neuronal cultures with CO2 overdose.

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