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Prepublished online as a Blood First Edition Paper on October 31, 2002; DOI 10.1182/blood-2002-07-1995.
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Blood, 15 April 2003, Vol. 101, No. 8, pp. 2905-2913
PLENARY PAPER
A highly sensitive strategy for SCID-repopulating cell assay by
direct injection of primitive human hematopoietic cells into NOD/SCID
mice bone marrow
Takashi Yahata,
Kiyoshi Ando,
Tadayuki Sato,
Hiroko Miyatake,
Yoshihiko Nakamura,
Yukari Muguruma,
Shunichi Kato, and
Tomomitsu Hotta
From the Division of Hematopoiesis, Research Center for
Regenerative Medicine, Department of Hematology, and Institute of
Medical Sciences, Tokai University School of Medicine, Isehara,
Kanagawa, Japan.
To measure the ability of human hematopoietic stem cells (HSCs),
the SCID-repopulating cell (SRC) assay has been widely used. Conventionally, human HSCs are transplanted into a nonobese
diabetic/severe combined immunodeficient (NOD/SCID) mouse via a tail
vein. However, those cells must go through various obstacles until they
reach the mouse marrow environment, which could explain the generally low homing efficiency in this system. Thus, the capability of HSCs may
not be studied accurately by this intravenous transplantation method. In our attempt to reveal actual SRC potential, ie, self-renewal and multilineage differentiation in recipient bone marrow, we introduced cells into mouse marrow directly (intrabone marrow [iBM])
to minimize the effect of factors that may interfere with the homing of
HSCs and compared the results obtained by intravenous and iBM methods.
When cord blood CD34+CD38 cells were
transplanted in NOD/SCID mice by iBM, a 15-fold higher frequency of
SRC, 1 in 44 CD34+CD38 cells, was achieved
compared with 1 in 660 by the intravenous method. Furthermore, the iBM
transplant showed high levels of engraftment in the secondary
transplantation. Pretreatment of CD34+ cells with
antibodies that block either very late antigen 4 (VLA-4) or
VLA-5 reduced engraftment partially, whereas blockage of both molecules
resulted in complete inhibition of engraftment, which suggests that
VLA-4 and VLA-5 are involved in different processes in engraftment or
have complementary roles. Our results indicate that the iBM injection
strategy is a more sensitive and direct way to measure the capability
of human SRCs and is useful to investigate the interaction of HSCs and
marrow environment in vivo.
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