A novel in vitro assay for murine haematopoietic stem cells.
Study of the biology of haematopoietic stem cells is crucially dependent on the availability of suitable in vitro assays. Existing assays have suffered from the fact that they detect small subcompartments of the total stem cell compartment. This limits experiments where it is required to assay a high proportion of stem cells, e.g. the enumeration of stem cell numbers under varying conditions or the identification and purification of stem cell regulators. We describe an in vitro assay which shows macroscopic colony formation and limited self-renewal capacity in vitro. The detected cell (CFU-A) has a low cycling status in normal bone marrow (NBM) and responds to known stem cell regulators. The incidence (100-200 per 10(5) in NBM), the proliferative characteristics under stress and some of the physical properties are similar to stem cells detected by colony formation after transplantation into lethally irradiated recipients (CFU-S). These data indicate that our assay detects a high proportion of haematopoietic stem cells in vitro. This will facilitate experiments on stem cell behaviour which have previously been difficult to conduct.
Eckmann
L
L
Beatson Institute for Cancer Research, Bearsden, Glasgow, UK.
Freshney
M
M
Wright
E G
EG
Sproul
A
A
Wilkie
N
N
Pragnell
I B
IB
D000818:Animals; D002455:Cell Division; D002469:Cell Separation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay / Q000379:methods*; D005434:Flow Cytometry; D005720:Gamma Rays; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051379:Mice; D013154:Spleen / Q000166:cytology / Q000528:radiation effects