Haematopoietic stem cells do not asymmetrically segregate chromosomes or retain BrdU.
Stem cells are proposed to segregate chromosomes asymmetrically during self-renewing divisions so that older ('immortal') DNA strands are retained in daughter stem cells whereas newly synthesized strands segregate to differentiating cells. Stem cells are also proposed to retain DNA labels, such as 5-bromo-2-deoxyuridine (BrdU), either because they segregate chromosomes asymmetrically or because they divide slowly. However, the purity of stem cells among BrdU-label-retaining cells has not been documented in any tissue, and the 'immortal strand hypothesis' has not been tested in a system with definitive stem cell markers. Here we tested these hypotheses in haematopoietic stem cells (HSCs), which can be highly purified using well characterized markers. We administered BrdU to newborn mice, mice treated with cyclophosphamide and granulocyte colony-stimulating factor, and normal adult mice for 4 to 10 days, followed by 70 days without BrdU. In each case, less than 6% of HSCs retained BrdU and less than 0.5% of all BrdU-retaining haematopoietic cells were HSCs, revealing that BrdU has poor specificity and poor sensitivity as an HSC marker. Sequential administration of 5-chloro-2-deoxyuridine and 5-iodo-2-deoxyuridine indicated that all HSCs segregate their chromosomes randomly. Division of individual HSCs in culture revealed no asymmetric segregation of the label. Thus, HSCs cannot be identified on the basis of BrdU-label retention and do not retain older DNA strands during division, indicating that these are not general properties of stem cells.
Kiel
Mark J
MJ
Howard Hughes Medical Institute, Life Sciences Institute, Department of Internal Medicine, and Centre for Stem Cell Biology, University of Michigan, Ann Arbor, Michigan 48109-2216, USA.
He
Shenghui
S
Ashkenazi
Rina
R
Gentry
Sara N
SN
Teta
Monica
M
Kushner
Jake A
JA
Jackson
Trachette L
TL
Morrison
Sean J
SJ
D000375:Aging; D000818:Animals; D000831:Animals, Newborn; D001854:Bone Marrow Cells / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000378:metabolism* / Q000494:pharmacology; D002478:Cells, Cultured; D020090:Chromosome Segregation / Q000187:drug effects; D003520:Cyclophosphamide / Q000494:pharmacology; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D013269:Stochastic Processes; D013997:Time Factors
D016179:Granulocyte Colony-Stimulating Factor; D003520:Cyclophosphamide; D001973:Bromodeoxyuridine