2 replies to this topic

[FACS_flow]

Hi there,

Does anyone of you know if the use of 10% DMSO, rather than glycerol, as cryoprotective agent,
in some way forces stem cells to undergo differentiation? So even if you would place the cells in
the freezing media, immediately freeze on dry ice and after thawing perform extensive washing
of thawed cells?
I am having some troubles after freezing primary cells isolated from tumor specimen as I
am seeing some weird results in the subsequent flow cytometry analyses...

Maybe someone knows an answer,

Cheers

[sameermahmood]

FACS_flow, on Dec 20 2009, 08:26 AM, said:

Hi there,

Does anyone of you know if the use of 10% DMSO, rather than glycerol, as cryoprotective agent,
in some way forces stem cells to undergo differentiation? So even if you would place the cells in
the freezing media, immediately freeze on dry ice and after thawing perform extensive washing
of thawed cells?
I am having some troubles after freezing primary cells isolated from tumor specimen as I
am seeing some weird results in the subsequent flow cytometry analyses...

Maybe someone knows an answer,

Cheers

I am working on ES cells and our lab uses 10% DMSO to freeze our cells. You can use the frozen vial for culture as long you the percent of differentiation do not exceed 5 - 10 %. If that is the case you have to go with a new one. Then why do you want to freeze them on dry ice. It just activate the differentiation process I think. You can place the cells in -80 C after suspending in freezing medium overnight and then transfer it to LN2 next day.
It should work.

Sameer

[DmitryM]

FACS_flow, on Dec 20 2009, 08:26 AM, said:

Hi there,

Does anyone of you know if the use of 10% DMSO, rather than glycerol, as cryoprotective agent,
in some way forces stem cells to undergo differentiation? So even if you would place the cells in
the freezing media, immediately freeze on dry ice and after thawing perform extensive washing
of thawed cells?
I am having some troubles after freezing primary cells isolated from tumor specimen as I
am seeing some weird results in the subsequent flow cytometry analyses...

Maybe someone knows an answer,

Cheers

Yes its a property of DMSO
its a some problem in reproductive cryopreservation. probably may use that experience? (cryoprotectors and proticols)