MSC from blood - (Aug/15/2007 )

Hai,
I have been working on islation of MSC from blood for the past few months.
I found out that there is always bacterias contamination occuring on the next few days of culture.
I am sure there is nothing wrong with my technique
Can someone please give me some advice on this problem and how to get blood as sterile as possible?


thanks in advance

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May I ask how do you obtain the blood?

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hai Minnie
I obtained the blood from patients using venupuncture technique
the needle is swabbed with 70% ethanol before punctured into the patient and the tubing draw the blood into an anti-coagulant container.
Later, the sample is transported back to lab, and isolation taking place under lamninar flow which is in sterile condition.

any wrong with my techniques?
any improvements/changes should i make? please advice

thanks in advance

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Hi,

Could you explain in detail how you isolate MSC and culture.
I do isolation of MSC from PB, BM as well as from fetal organ, but I never got contamination so, I am kinda curious.

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I extract WBC from boffy coat …..and always I keep 2 points in mind
First …..in transferring the blood from the bag through the pipe to the container …..the pipe has to be spread by 70% alcohol and then left to dry. making sure the it does not touch anything ….second is the bag should not touch the tip of the container…..

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hi..im new to this forum..basically im doing the same method as yours..and i also encounter the same problem..bacteria contamination..though it dont really look like one...at first the cells are ok..but then contamination strike out in our lab..and i havent recover from the problem since August last year..the media is clear but u can see with ur naked eyes some kind of debris in the middle of the well (i use 24 well plate)..cells are also healthy..do u guys have any idea what is the problem that im facing right now? plz help

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