PMID- 2333304
OWN - NLM
STAT- MEDLINE
DCOM- 19900606
LR  - 20190501
IS  - 0027-8424 (Print)
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Linking)
VI  - 87
IP  - 9
DP  - 1990 May
TI  - Functional characterization of individual human hematopoietic stem cells cultured 
      at limiting dilution on supportive marrow stromal layers.
PG  - 3584-8
AB  - A major goal of current hematopoiesis research is to develop in vitro methods 
      suitable for the measurement and characterization of stem cells with long-term in 
      vivo repopulating potential. Previous studies from several centers have suggested 
      the presence in normal human or murine marrow of a population of very primitive 
      cells that are biologically, physically, and pharmacologically different from 
      cells detectable by short-term colony assays and that can give rise to the latter 
      in long-term cultures (LTCs) containing a competent stromal cell layer. In this 
      report, we show that such cultures can be used to provide a quantitative assay 
      for human "LTC-initiating cells" based on an assessment of the number of 
      clonogenic cells present after 5-8 weeks. Production of derivative clonogenic 
      cells is shown to be absolutely dependent on the presence of a stromal cell 
      feeder. When this requirement is met, the clonogenic cell output (determined by 
      assessment of 5-week-old cultures) is linearly related to the input cell number 
      over a wide range of cell concentrations. Using limiting dilution analysis 
      techniques, we have established the frequency of LTC-initiating cells in normal 
      human marrow to be approximately 1 per 2 X 10(4) cells and in a highly purified 
      CD34-positive subpopulation to be approximately 1 per 50-100 cells. The 
      proliferative capacity exhibited by individual LTC-initiating cells cultured 
      under apparently identical culture conditions was found to be highly variable. 
      Values for the number of clonogenic cells per LTC-initiating cell in 5-week-old 
      cultures ranged from 1 to 30 (the average being 4) with similar levels being 
      detected in positive 8-week-old cultures. Some LTC-initiating cells are 
      multipotent as evidenced by their generation of erythroid as well as 
      granulopoietic progeny. The availability of a system for quantitative analysis of 
      the proliferative and differentiative behavior of this newly defined compartment 
      of primitive human hematopoietic cells should facilitate future studies of 
      specific genetic or microenvironmental parameters involved in the regulation of 
      these cells.
FAU - Sutherland, H J
AU  - Sutherland HJ
AD  - Terry Fox Laboratory, Cancer Control Agency, British Columbia, Vancouver, Canada.
FAU - Lansdorp, P M
AU  - Lansdorp PM
FAU - Henkelman, D H
AU  - Henkelman DH
FAU - Eaves, A C
AU  - Eaves AC
FAU - Eaves, C J
AU  - Eaves CJ
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
SB  - IM
MH  - *Bone Marrow Cells
MH  - Cell Count
MH  - Cell Separation
MH  - Cells, Cultured
MH  - Colony-Forming Units Assay
MH  - Flow Cytometry
MH  - Hematopoietic Stem Cells/*cytology
MH  - Humans
MH  - Time Factors
PMC - PMC53946
EDAT- 1990/05/01 00:00
MHDA- 1990/05/01 00:01
PMCR- 1990/11/01
CRDT- 1990/05/01 00:00
PHST- 1990/05/01 00:00 [pubmed]
PHST- 1990/05/01 00:01 [medline]
PHST- 1990/05/01 00:00 [entrez]
PHST- 1990/11/01 00:00 [pmc-release]
AID - 10.1073/pnas.87.9.3584 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 1990 May;87(9):3584-8. doi: 10.1073/pnas.87.9.3584.