Limiting factors in murine hematopoietic stem cell assays

by Alexey Bersenev on November 26, 2009 · 0 comments

in hematopoietic, reviews

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All of papers in “reviews” category is kindly picked by us and highly recommended to read. We will try to pick reviews mostly in open access.

I’m happy to share one of my favorite methodological reviews – Limiting factors in murine hematopoietic stem cell assays by David Scadden and Louise Purton, published in Cell Stem Cell in 2007.

This great review is briefly summarizing all assays that we use to study hematopoietic stem cells. What I especially like is that how authors highlight challenges, limitations and controversies in hematopoietic stem cells assays.

Some questions about the “gold standard” – bone marrow transplantation assays discussed in review:

Competing cells (whole BM versus progenitors (c-Kit+ or Sca-1+) depleted, cell number)

Donor (test) cells (whole BM versus purified progenitors or HSC populations; freshly isolated vs cultured HSC )

Donor (test) cells from single mouse or pulled from few mice

Donor cell % and lineages used to determine the number of negative/positive mice (1% versus 0.1%; multilineage versus any 1 lineage)

Time posttransplant at which the results are analyzed (3 or 6 months)

Method of detecting donor vs host cells (CD45.1-CD45.2; Y-chromosome, GFP)

Recipient conditioning (lethal versus non-irradiated versus sublethal)

In conclusion, while remaining one of the better assays to assess HSC content, investigators should also be aware of the potential problems associated with analyzing limiting dilution assays. Caution should also be taken when forming conclusions based on data obtained using shorter-term approaches commonly used to analyze HSC content. It is recommended that investigators assessing HSC content in mutant and nonsteady state populations use long-term in vivo-repopulating assays together with a range of the other short-term in vitro and in vivo assays to accurately determine the number and functional potential of HSCs and their progenitors in mice. Furthermore, the recommended 16–26 weeks posttransplant required for analysis have cost implications that are obvious and extremely problematic. Finally, the long interval required for analysis of these assays impedes progress in the field. Alternative means of assessing and quantitating HSCs are clearly needed but may be more of a dream than a reality.

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