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Phenotype and function of human hematopoietic cells engrafting immune-
deficient CB17-severe combined immunodeficiency mice and nonobese
diabetic-severe combined immunodeficiency mice after transplantation of
human cord blood mononuclear cells
F Pflumio, B Izac, A Katz, LD Shultz, W Vainchenker and L Coulombel
INSERM U362, Institut Gustave Roussy, Villejuif, France.
In an attempt to understand better the regulation of stem cell function in
chimeric immunodeficient mice transplanted with human cells, and the
filiation between progenitor cells identified in vitro and in vivo, we
assessed the different compartments of hematopoietic progenitors found in
the marrow of CB17-severe combined immunodeficiency (SCID) mice (34 mice, 9
experiments) after intravenous injection of 2 to 3 x 10(7) cord blood
mononuclear cells. On average 6.3 +/- 4 x 10(5) human cells were detected
per four long bones 4 to 6 weeks after the transplant predominantly
represented by granulomonocytic (CD11b+) and B lymphoid (CD19+) cells.
Twenty five percent of these human cells expressed the CD34 antigen, of
which 90% coexpressed the CD38 antigen and 50% the CD19 antigen. Functional
assessment of progenitor cells (both clonogenic and long-term
culture-initiating cells [LTC-IC]) was performed after human CD34+ cells
and CD34+/CD38- cells have been sorted from chimeric CB17-SCID marrow 3 to
10 weeks after intravenous (IV) injection of human cells. The frequency of
both colony-forming cells and LTC-IC was low (4% and 0.4%, respectively in
the CD34+ fraction) when compared with the frequencies of cells with
similar function in CD34+ cells from the starting cord blood mononuclear
cells (26% +/- 7% and 7.2% +/- 5%, respectively). More surprisingly, the
frequency of LTC-IC was also low in the human CD34+ CD38- fraction sorted
from chimeric mice. This observation might be partly accounted for by the
expansion of the CD34+ CD19+ B-cell precursor compartment. Despite their
decreased frequency and absolute numbers, the differentiation capability of
these LTC-IC, assessed by their clonogenic progeny output after 5 weeks in
coculture with murine stromal cells was intact when compared with that of
input LTC-IC. Furthermore the ratio between clonogenic progenitor cells and
LTC-IC was similar in severe combined immunodeficiency (SCID) mice studied
4 weeks after transplant and in adult marrow or cord blood suspensions.
Results generated in experiments where nonobese diabetic (NOD)-SCID mice
were used as recipients indicate a higher level of engraftment but no
change in the distribution of clonogenic cells or LTC-IC. These results
suggest that the hierarchy of hematopoietic differentiation classically
defined in human hematopoietic tissues can be reconstituted in
immunodeficient SCID or NOD-SCID mice.
Volume 88,
Issue 10,
pp. 3731-3740,
11/15/1996
Copyright © 1996 by The American Society of Hematology
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