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BRIEF REPORT
From the University of Minnesota Cancer Center and
Department of Pediatrics, Division of Bone Marrow Transplantation,
Minneapolis; and The Jackson Laboratories, Bar Harbor, ME.
In utero transplantation (IUT) is becoming a viable option for the
treatment of various immune and metabolic disorders diagnosed early in
gestation. In this study, donor fetal liver cells had a 10-fold
competitive engraftment advantage relative to adult bone marrow in
allogeneic fetal severe combined immunodeficient (SCID) recipients
compared with adult recipients. In contrast, adult bone marrow cells
engrafted slightly better than fetal liver cells in allogeneic adult
SCID transplant recipients. By using different ratios of fetal and
adult cell mixtures, fetal liver cells repopulated 8.2 times better
than adult bone marrow cells in fetal recipients, but only 0.8 times as
well in adult recipients. Fetal SCID recipients were more permissive to
an allogeneic donor graft than adult recipients. These data indicate
that the recipient microenvironment may regulate the engraftment
efficiency of a given stem cell source and suggest that the use of cord
blood should be tested in clinical IUT.
(Blood. 2002;99:1870-1872) In utero transplantation (IUT) offers a therapeutic
option for the treatment of hemoglobinopathies and immune or metabolic disorders diagnosed early in gestation.1-8 The preimmune
fetal recipient may be more tolerant than an adult recipient to an
unmatched donor stem cell source in the absence of potential toxicities associated with conventional transplantation-conditioning
regimes.6,9-11 Importantly, early successful establishment
of donor stem cell engraftment prenatally may prevent or reduce the
pathology associated with the underlying disorder. Fetal liver (FL),
cord blood, and adult bone marrow (BM) are potential stem cell sources
for transplantation. The relative engraftment efficiency of each source
may depend on intrinsic differences in cytokine requirements as well as
extrinsic signals that differ in the fetal versus the adult
microenvironment.11,12 In this study, we compare the
relative engraftment capacity of allogeneic FL versus adult BM in both
fetal and adult severe combined immunodeficient (SCID) recipients.
Mice
IUT and adult transplantation
Flow cytometry Peripheral blood leukocytes (PBLs) were stained with fluorescein isothiocyanate-, phycoerythrin-, or biotin-conjugated antibodies to CD4, CD8, MAC-1 (CD11b), CD19, CD45.1, CD45.2, H2b, and H2d with SA-peridinin chlorophyll protein (PharMingen, San Diego, CA) and analyzed on a FACScalibur by using CellQuest software (Becton Dickinson, San Jose, CA).
To determine the relative engraftment capacity of FL versus
T-cell-depleted adult BM in fetal recipients, BALB/c SCID fetuses were
injected intraperitoneally on day 15/16 of gestation with equal numbers
of C57BL/6 CD45.2 adult T-cell-depleted BM and C57BL/6 CD45.1 FL
cells. PBLs were typed for donor H2Kb and CD45.1 and CD45.2
at 2 months after birth. Thirty-four (94%) of 36 IUT recipients had
evidence of donor engraftment (
Although long-term reconstituting stem cells have been reported to be approximately 7 times more frequent in FL at 12 to 15 days of gestation than in adult BM, the frequency has been reported to decline approximately 3-fold by 15.5 days of gestation.14,15 Because FL was obtained at 15 to 16 days of gestation, we hypothesized that the striking competitive engraftment advantage of FL as compared with adult BM in fetal recipients may not be due solely to an increased stem cell content in FL but may be due also to a favorable fetal microenvironment. To compare engraftment of FL cells to adult BM in fetal versus
unconditioned adult SCID recipients, ratios of 1:1, 0.3:1, and 0.1:1 FL
to adult BM were administered (Table 2).
Fetal and adult transplant recipients received an approximately
equivalent cell dose on a per-weight basis (range,
120-1200 × 106 cells/kg depending on FL/BM ratio). These
formal dose response studies indicated FL had a substantial competitive
engraftment advantage over adult BM in IUT recipients. Mice that
underwent transplantation as fetuses with a 1:1 ratio of FL to
adult BM had 81% of PBLs originating from donor FL and 10%
originating from donor adult BM. Reducing the ratio of FL to adult BM
to 0.1:1 resulted in 42% and 46% of cells originating from donor FL
and donor adult BM, respectively. Repopulating units (RUs) were
calculated for FL, where 1 RU was defined as equal to the repopulating
ability of 105 adult BM cells.16 FL produced
RU numbers directly proportional to FL numbers used over a 10-fold
range (Figure 1) and averaged 8.2 RU per
105 FL; thus, FL repopulated 8.2 times as well as BM in
allogeneic fetal SCID recipients.
In contrast to these data in IUT recipients, FL and adult BM had an approximate equivalent capacity to reconstitute an adult recipient. Five months after transplantation, adult transplant recipients of a 1:1 ratio of FL to adult BM had 36% and 39% of PBLs originating from donor FL and donor adult BM, respectively (Table 2). Reducing the ratio of FL to adult BM to 0.1:1 in adult recipients resulted in only 1% to 3% of cells originating from the donor FL stem cell source. FL produced RU values proportional to FL numbers used (Figure 1) but only averaged 0.8 RU per 105 FL; thus, FL repopulated only 0.8 times as well as BM in allogeneic adult SCID recipients. These results in adult mice contrast with earlier studies in which fetal cells had a 3.5- to 7.0-fold long-term engraftment advantage as compared with adult cells in adult recipients.17 However, those studies used lethally irradiated syngeneic adult recipients rather than nonirradiated allogeneic SCID recipients used in the current study, suggesting that the host conditioning regime or allogenicity of the donor may be important factors. Furthermore, no previous study has compared fetal and adult cells in both fetal and adult recipients. Additionally, these data indicate that fetal SCID recipients are more readily engraftable with an allogeneic donor graft than are adult SCID recipients. In contrast to the high engraftment rate (90%) in fetal IUT recipients, only 10 (53%) of 19 adult transplant recipients had evidence of donor engraftment when typed at 6 to 7 weeks after transplantation. Mice that underwent transplantation as fetuses also had higher levels of donor engraftment than adult transplant recipients. Collectively, these data indicate that fetal recipients are more permissive to an allogeneic donor graft than are adult recipients and further that the recipient microenvironment may regulate the proliferative responses of a given stem cell source. FL had an 8.2-fold competitive engraftment advantage as compared with adult BM in fetal recipients. In contrast, FL engrafted only 0.8 times as well as the adult BM in adult transplant recipients. Our data may have important clinical implications and suggest that fetal hematopoietic cells (eg, cord blood cells) may be preferable to adult BM for IUT in humans.
Submitted July 12, 2001; accepted October 26, 2001.
Supported by grants R01 HL49997 and R01 HL52952 (B.R.B.) as well as R01 HL63230 and R01 HL58820 (D.E.H.) from the National Institutes of Health.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Bruce R. Blazar, MMC 109, University of Minnesota Hospital, Minneapolis, MN 55455; e-mail: blaza001{at}tc.umn.edu.
1.
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Adult bone marrow-derived pluripotent hematopoietic stem cells are engraftable when transferred in utero into moderately anemic fetal recipients.
Blood.
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In utero transfer of adult bone marrow cells into recipients with severe combined immunodeficiency disorder yields lymphoid progeny with T- and B-cell functional capabilities.
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In utero transplantation of fetal liver cells in the mucopolysaccharidosis type VII mouse results in low-level chimerism, but overexpression of beta-glucuronidase can delay onset of clinical signs.
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Flake AW, Roncarolo MG, Puck JM, et al.
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1996;335:1806-1810 5. Flake AW, Zanjani ED. In utero hematopoietic stem cell transplantation. A status report. JAMA. 1997;278:932-967[Abstract]. 6. Jones DR, Bui TH, Anderson EM, et al. In utero haematopoietic stem cell transplantation: current perspectives and future potential. Bone Marrow Transplant. 1996;18:831-867[Medline] [Order article via Infotrieve]. 7. Touraine JL, Raudrant D, Royo C, et al. In utero transplantation of hemopoietic stem cells in humans. Transplant Proc. 1991;23:1706-1708[Medline] [Order article via Infotrieve]. 8. Wengler GS, Lanfranchi A, Frusca T, et al. In-utero transplantation of parental CD34 haematopoietic progenitor cells in a patient with X-linked severe combined immunodeficiency (SCIDXI). Lancet. 1996;348:1484-1487[CrossRef][Medline] [Order article via Infotrieve]. 9. Zanjani ED, Pallavicini MG, Ascensao JL, et al. Engraftment and long-term expression of human fetal hemopoietic stem cells in sheep following transplantation in utero. J Clin Invest. 1992;89:1178-1188.
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© 2002 by The American Society of Hematology.
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  J. Chan, S. N. Waddington, K. O'Donoghue, H. Kurata, P. V. Guillot, C. Gotherstrom, M. Themis, J. E. Morgan, and N. M. Fisk Widespread Distribution and Muscle Differentiation of Human Fetal Mesenchymal Stem Cells After Intrauterine Transplantation in Dystrophic mdx Mouse Stem Cells, April 1, 2007; 25(4): 875 - 884. [Abstract] [Full Text] [PDF]
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P. V. Guillot, C. Gotherstrom, J. Chan, H. Kurata, and N. M. Fisk Human First-Trimester Fetal MSC Express Pluripotency Markers and Grow Faster and Have Longer Telomeres Than Adult MSC Stem Cells, March 1, 2007; 25(3): 646 - 654. [Abstract] [Full Text] [PDF]
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 M. A. Suckow, A. Zollman, I. Cornelissen, M. Casad, J. Roahrig, F. J. Castellino, and E. D. Rosen Tissue Distribution of Fetal Liver Cells Following In Utero Transplantation in Mice Experimental Biology and Medicine, December 1, 2005; 230(11): 860 - 864. [Abstract] [Full Text] [PDF]
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| Copyright © 2002 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||||
Top
Abstract
Introduction
2% donor cells). FL engrafted
approximately 10-fold better than adult BM (Table
BALB/c SCID)
reconstituted lethally irradiated adult C57BL/6 recipients with both
C57BL/6 CD45.1 (donor FL origin) and CD45.2 (donor BM origin) cells. In
these secondary recipients, as in the original IUT recipients, there
were 8 to 10 times more cells from donor CD45.1 FL than from adult BM
origin (data not shown). Collectively, these data indicate that fetal
hematopoietic cells may be preferable to adult hematopoietic cells
for IUT.
