Should i use cells healed from mycoplasma - (Dec/29/2008 )
Hey guys
I have 2 questions :
I have SHSY5Y cells (neuroblastoma cell line) that was treated for mycoplasma contamination and do not have it anymore .
The problem is that they stopped to react to glutamate, stopped dying from that and they were really sensetive to that before .
Could that happen because of mycoplasma? Also should i use those cells at all even if they were healed from the mycoplasma?
What do you think
Also the youngiest passage that i have is 18 ? The atcc say you can use it only up to 20 passage? Do you use it above 20 passage?
How do you define passage?
thanks , Hope you can help me
-Emily83-
QUOTE (Emily83 @ Dec 29 2008, 12:42 PM)
Hey guys
I have 2 questions :
I have SHSY5Y cells (neuroblastoma cell line) that was treated for mycoplasma contamination and do not have it anymore .
The problem is that they stopped to react to glutamate, stopped dying from that and they were really sensetive to that before .
Could that happen because of mycoplasma? Also should i use those cells at all even if they were healed from the mycoplasma?
What do you think
Also the youngiest passage that i have is 18 ? The atcc say you can use it only up to 20 passage? Do you use it above 20 passage?
How do you define passage?
thanks , Hope you can help me
I have 2 questions :
I have SHSY5Y cells (neuroblastoma cell line) that was treated for mycoplasma contamination and do not have it anymore .
The problem is that they stopped to react to glutamate, stopped dying from that and they were really sensetive to that before .
Could that happen because of mycoplasma? Also should i use those cells at all even if they were healed from the mycoplasma?
What do you think
Also the youngiest passage that i have is 18 ? The atcc say you can use it only up to 20 passage? Do you use it above 20 passage?
How do you define passage?
thanks , Hope you can help me
ANSWER TO FIRST QUESTION..... " IF IN DOUBT, CHUCK THEM OUT"
ANSWER TO SECOND QUESTION.....THE ATCC ARE WORLD EXPERTS IN TISSUE CULTURE AND GIVE BEST ADVICE........WHY QUESTION THEM........BUY MORE CELLS
You have really answered your own question though...think about it!!!!!!!
"The problem is that they stopped to react to glutamate, stopped dying from that and they were really sensetive to that before"
As I always say to any researcher who will listen, cleaning up mycoplasma infection will not revert the cells back to their original phenotype.......the cells and their characteristics maybe changed forever.
If the cells are commercially available them buy them i.e. from the ATCC/ECACC.
DO NOT TRY AND SAVE MONEY AND OBTAIN THEM FROM ANOTHER LAB NEXT OR A COLLABORATOR. You do not know whether they have:
Regularly mycoplasma tested these cells.
Kept them in the right conditions
Are they the passage number you require
......The list of variables is endless................BUY SOME MORE.
Please listen to Uncle Rhombus....he knows best........the advice above comes from 30 years of experience with cells/tissues in culture
Hope this is useful......HAPPY NEW YEAR
rHOMBUS
-Rhombus-
QUOTE (Rhombus @ Jan 2 2009, 07:24 AM)
QUOTE (Emily83 @ Dec 29 2008, 12:42 PM)
Hey guys
I have 2 questions :
I have SHSY5Y cells (neuroblastoma cell line) that was treated for mycoplasma contamination and do not have it anymore .
The problem is that they stopped to react to glutamate, stopped dying from that and they were really sensetive to that before .
Could that happen because of mycoplasma? Also should i use those cells at all even if they were healed from the mycoplasma?
What do you think
Also the youngiest passage that i have is 18 ? The atcc say you can use it only up to 20 passage? Do you use it above 20 passage?
How do you define passage?
thanks , Hope you can help me
I have 2 questions :
I have SHSY5Y cells (neuroblastoma cell line) that was treated for mycoplasma contamination and do not have it anymore .
The problem is that they stopped to react to glutamate, stopped dying from that and they were really sensetive to that before .
Could that happen because of mycoplasma? Also should i use those cells at all even if they were healed from the mycoplasma?
What do you think
Also the youngiest passage that i have is 18 ? The atcc say you can use it only up to 20 passage? Do you use it above 20 passage?
How do you define passage?
thanks , Hope you can help me
ANSWER TO FIRST QUESTION..... " IF IN DOUBT, CHUCK THEM OUT"
ANSWER TO SECOND QUESTION.....THE ATCC ARE WORLD EXPERTS IN TISSUE CULTURE AND GIVE BEST ADVICE........WHY QUESTION THEM........BUY MORE CELLS
You have really answered your own question though...think about it!!!!!!!
"The problem is that they stopped to react to glutamate, stopped dying from that and they were really sensetive to that before"
As I always say to any researcher who will listen, cleaning up mycoplasma infection will not revert the cells back to their original phenotype.......the cells and their characteristics maybe changed forever.
If the cells are commercially available them buy them i.e. from the ATCC/ECACC.
DO NOT TRY AND SAVE MONEY AND OBTAIN THEM FROM ANOTHER LAB NEXT OR A COLLABORATOR. You do not know whether they have:
Regularly mycoplasma tested these cells.
Kept them in the right conditions
Are they the passage number you require
......The list of variables is endless................BUY SOME MORE.
Please listen to Uncle Rhombus....he knows best........the advice above comes from 30 years of experience with cells/tissues in culture
Hope this is useful......HAPPY NEW YEAR
rHOMBUS
hi
I fairly agree with Rhombus.
I have experienced contamination with mycoplasma, and as same you i turned to this amazing forum. mostly users adviced me to throw everything out and start over. i tried to heal some cells with Bm cyclin but i could see that my cells have been not the same. i work with immune responses in viral infections and i noticed that viral infection and theirs levels were drastically lower than before, and high levels of interferons with no viral infection( i'm not sure if mycoplasmas can trigger interferon pathways but i have related this)
my advice. Throw everything aout and start over again!!
Best
Reis,V.P
-Reis V.P-