5 replies to this topic

[sssss]

hi,
how to store haematopoietic stem cells for long term use????

[si78to]

sssss, on Oct 7 2009, 04:45 AM, said:

hi,
how to store haematopoietic stem cells for long term use????

I am working with HSC isolated from cord blood. I cryofreezed at -152C using DMSO. But usually some of the cell will die after revived.

[boudou_611]

si78to, on Nov 11 2009, 03:32 PM, said:

sssss, on Oct 7 2009, 04:45 AM, said:

hi,
how to store haematopoietic stem cells for long term use????

I am working with HSC isolated from cord blood. I cryofreezed at -152C using DMSO. But usually some of the cell will die after revived.

Hi,

I cryopreserved HSC in freezing media: 60% FBS, 10% DMSO & 30% culture media. after cryopreserved and thawing, the cells viability will be low at initial but then they will recover..

Rgrds,
Boudou

[Hodag]

si78to, on Nov 11 2009, 03:32 PM, said:

sssss, on Oct 7 2009, 04:45 AM, said:

hi,
how to store haematopoietic stem cells for long term use????

This is very tough to do, mainly because you probably don't have a single cell type but rather a "cell soup" containing stem cells, progenitor, and some precursors.  All these cells have different diameters and it is almost impossible to get a uniform freeze/thaw when working with a mixed cell population so a significant amount of lysis is practically guaranteed.

If you have one particlar phenotype that you are interested in, you may want to optimize the process to preserve that cell.

[sssss]

hiiiiiiii
thanksa a lot for your suggestions
but how to store a lineage negative infected cells?????????

[Piersgb]

Just out of interest, where do you source your haematopoietic stem cells from and what are you doing with them?

I think as people previously said you have to tailor your freezing method/protocol specifically to your stage of culture or the cell type that you have isolated. If it is still a mixture then there is sure to be a reasonable loss of cell viability after thawing.

I'm currently sourcing mouse bone marrow cells from the long bones of mice, putting these unselected cells into culture to eventually produce eosinophils. So far with little sucess! Have been following a published protocol tho...



"Functionally competent eosinophils differentiated ex vivo in high purity from normal mouse bone marrow", J Immunol. 2008 Sep 15;181(6):4004-9.



Anyone else doing anything similar?!

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