Spectra/Por, the only name in laboratory dialysis.

CellCultureChat.com:   The Answers for All of Your Cell Culture Questions.


Close MessageUserDate(yyyy-mm-dd)
Previous Subject
Next Subject 		contamination before freezingdorarat2008-12-04Click here to register.
Hi! As a first time student in cell culture course, I made a mistake of taking out the whole culture from laf bench out to the non sterile environment and took a ul of that using non sterile pipette before freezing the culture. So it is already contaminated, what will be the outcome and solutions?
 
Re: contamination before freezing
lubrol2008-12-05Click here to register.
The outcome would be the loss of the entire frozen culture if it is not "cured" after thawing. The contamination will easily overgrow the culture.
As to a "cure", that is a bit trickier. It will depend on the nature of the contaminant and how robust the culture itself is. Assuming a valuable culture and one that is reasonably robust: if the culture is not very valuable and especially if it is easily replaced; replacing it is the better option. If not:
The best bet is to gently spin down the freshly thawed culture and rinsing the pellet with sterile PBS 2 - 3 times. After the last rinse, plate the cell in the regular medium containing pen-strep-fungizone (p-s-f) (this is available as a 100X concentrate from most cell culture media companies). Change the medium after 24 hours and again after 72 hours. If the cells look healthy after that, allow them to grow to confluency and then split the culture as normal EXCEPT plate half of the cells into medium with the p-s-f and half without. let them grow as you normally would and examine the culture without the antibiotics fir bacterial of fungal growth.

In a way you are lucky that you immediately frozen the cultures. That minimized the chances of the contamination over-growing the culture.


Hope this helps,

Lubrol
 
Re: contamination before freezing
dorarat2008-12-06Click here to register.
Thanx so much!

Thanks for visiting.
  • To ask or answer a question, you need to register here.
  • You can browse by clicking on a message subject to see the complete message.
  • If you have previously registered, you can log in below.

Message (click on a subject to view the messages)
Click for older messages
OlderOlder LastOldest
Click for newer messages
NewestFirst LastNewer
Search for messages that contain  

You must register before asking a question or providing an answer.
You can click here to register. If you have already registered you can log in below.
(If you check the Use Cookies box then your computer will remember your log in name.)
User Name:
Password:


Click here if you previously registered and remember your user name but not your password.
Email us at cccc@lplc.com if you are having other problems.


Back to Top