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Blood, Vol. 95 No. 9 (May 1), 2000:
pp. 2813-2820
Isolation and characterization of human
CD34 Lin and
CD34+Lin hematopoietic stem cells using
cell surface markers AC133 and CD7
Lisa Gallacher,
Barbara Murdoch,
Dongmei M. Wu,
Francis N. Karanu,
Mike Keeney, and
Mickie Bhatia
From The John P. Robarts Research Institute, London,
Ontario, Canada, and the Department of Microbiology and Immunology,
University of Western Ontario, London, Ontario, Canada.
Recent evidence indicates that human hematopoietic stem cell
properties can be found among cells lacking CD34 and lineage commitment markers (CD34 Lin). A
major barrier in the further characterization of human
CD34 stem cells is the inability to detect this
population using in vitro assays because these cells only
demonstrate hematopoietic activity in vivo. Using cell surface
markers AC133 and CD7, subfractions were isolated within
CD34CD38Lin and
CD34+CD38Lin cells derived
from human cord blood. Although the majority of CD34CD38Lin cells
lack AC133 and express CD7, an extremely rare population of
AC133+CD7 cells was identified at a
frequency of 0.2%. Surprisingly, these AC133+CD7 cells were highly enriched for
progenitor activity at a frequency equivalent to purified fractions of
CD34+ stem cells, and they were the only subset among the
CD34CD38Lin population
capable of giving rise to CD34+ cells in defined liquid
cultures. Human cells were detected in the bone marrow of
non-obese/severe combined immunodeficiency (NOD/SCID) mice 8 weeks
after transplantation of ex vivo-cultured AC133+CD7 cells isolated from the
CD34CD38Lin population,
whereas 400-fold greater numbers of the
AC133CD7 subset had no engraftment
ability. These studies provide novel insights into the hierarchical
relationship of the human stem cell compartment by identifying a rare
population of primitive human CD34 cells that are
detectable after transplantation in vivo, enriched for in vitro
clonogenic capacity, and capable of differentiation into
CD34+ cells.
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