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HEMATOPOIESIS
From the Division of Hematology/Oncology and Indiana
Elks Cancer Research Center, Department of Medicine; Herman B Wells
Center for Pediatric Research, Department of Pediatrics; and Department
of Microbiology and Immunology, Indiana University School of Medicine,
Indianapolis.
Engraftment potential of hematopoietic stem cells (HSCs) is likely
to be dependent on several factors including expression of
certain adhesion molecules (AMs) and degree of mitotic quiescence. The
authors investigated the functional properties and engraftment potential of Sca-1+lin The ability of hematopoietic stem cells (HSCs) to
reconstitute normal bone marrow (BM) hematopoiesis following
transplantation into suitable recipients relies on the potential of
these cells to home to and anchor within the BM microenvironment.
Homing is an intricate process by which HSCs, through interactions
between adhesion molecules (AMs) and their counter-receptors expressed on BM endothelium, migrate through endothelial cells and into the
stromal cell microenvironment. Within the BM microenvironment, the
homing process continues as HSCs, again through interactions between
AMs and cognant ligands, specifically anchor within appropriate BM
niches and begin the process of hematopoiesis. Mobilization of HSCs
from their BM niches into the periphery following administration of
growth factors or chemotherapeutic agents is likely to involve sequential loss or alterations in adhesive interactions between HSCs
and stromal cells, and then endothelium, with final release into the
periphery. Although data implicating certain AMs in various stages of
stem cell homing and egress from the BM are beginning to
accumulate,1-7 much remains to be learned regarding the
relationship between AMs and stem cell trafficking, and how this
process affects stem cell engraftment and long-term hematopoiesis in
transplanted recipients.
While AMs are likely to direct the trafficking of HSCs within the BM
microenvironment and periphery, the position of these cells in specific
phases of cell cycle is believed to dictate the hematopoietic potential
of HSCs.8 A large body of evidence in both in vitro and in
vivo systems supports the notion that mitotic quiescence is a
fundamental characteristic of HSCs and is essential for the
preservation of primitive hematopoietic function. Recent reports in the
murine,9 feline,10 and human11
systems suggest that the cell cycle of HSCs may be relatively
long and that these cells may display reduced engraftment potential at the time of active cell division.12-14 On the other hand,
few reports seem to indicate that cells capable of long-term
engraftment cycle quickly following transplantation.15
These data pose the question of whether changes in AM status,
concurrent with entry of these cells into active cell division,
influence the engraftment potential of cycling HSCs. We and others have
begun to investigate relationships between cell cycle progression and
expression or function of AMs on primitive hematopoietic progenitor
cells (HPCs). Some reports recently demonstrated an increased
expression of certain AMs16-19 and, in some cases,
increased adhesion17 of primitive HPCs in active phases of
cell cycle. These data, which clearly establish a link between cell
cycle status and AM repertoire, suggest the possible existence of
regulatory control mechanisms between expression or function of AMs and
cell cycle position of HSCs, which may in turn have an impact on the
engraftment potential of these cells.
In the present study, we examined the contribution of 6 AMs to
the homing and long-term engraftment potential of primitive HPCs using
an in vivo murine BM transplantation model. Results of these studies
imply a theoretical phenotype of a Sca-1+lin Mice
Flow cytometric cell sorting and analysis of
Sca-1+lin
To isolate Sca-1±lin To analyze the dual expression of AM on
Sca-1+lin Separation of cell cycle subfractions of
Sca-1+lin  AM+ or
AM cells enriched for engraftment potential were isolated
as described above, resuspended in Hoechst buffer (Hanks' balanced
salt solution [Biowhittaker, Walkersville, MD], 20 mmol/L
HEPES [Biowhittaker], 1 g/L glucose, and 10% fetal calf serum [FCS]), and stained with Hoechst 33342 (Molecular Probes) at 10 µmol/L for 45 minutes at 37°C as previously
described.21 To limit dye efflux via the MDR-1
pump, 100µmol/L verapamil (Sigma Chemical Co, St Louis, MO) was added
to the staining buffer.22 Cells were washed and
resuspended in Hoechst buffer plus verapamil for cell sorting. Cells
falling within a primary light scatter gate and containing 2n DNA
were gated and sorted as G0/G1. Cells containing between 2n and 4n DNA were gated and sorted as
S/G2+M. Hypodiploid events, when present, were excluded
from sort windows. When quantities were sufficient, a small portion of
sorted cells were subjected to postsort analyses, either by examining
Hoechst fluorescence or by propidium iodide staining, as previously
described.23
HPP-CFC and LPP-CFC assay Between 0.8 × 103 and 1.5 × 103 Sca-1+lin cells or AM subfractions were
suspended in triplicate in 1 mL double-layer agar cultures and assayed
for HPP-CFCs and LPP-CFCs as previously described.24 Cultures were incubated in a 100%-humidified 5% O2, 10%
CO2, and 85% N2 environment. Recombinant
hematopoietic growth factors were used as follows: 200 U/mL murine
interleukin-3 (mIL3), 1000 U/mL mIL1- , 50 ng/mL murine stem cell
factor (mSCF), 25 ng/mL murine granulocyte macrophage-colony
stimulating factor (mGM-CSF) (all from PeproTech, Rocky Hill, NJ), and
1600 U/mL human macrophage-CSF-1 (hM-CSF, Genetics Institute, Camden,
MA). On day 14, colonies larger than 0.5 mm were scored as HPP-CFC and
those smaller than 0.5mm as LPP-CFC.
Transplantation protocol C57BL/6 female recipients between 10 and 12 weeks of age were lethally irradiated (split dose of 700 centrigrays [cGy] followed by 350 cGy 3 to 4 hours later) from a 137Cs gamma irradiator (GammaCell 40; Nordion International, Kanata, Ontario, Canada). Mice were transplanted via tail vein injections 3 to 6 hours later with 0.25 × 103 to 1.0 × 103 donor B6.Gpi-1a or B6.BoyJ cells, along with 0.3 × 105 to 1.0 × 105 competitor cells (low-density BM cells of C57BL/6 origin), as described in Figures. Recipient mice were bled from the tail vein monthly until 6 or 8 months posttransplantation for analysis of donor-derived hematopoiesis.Determination of donor chimerism in transplanted recipients Chimerism in C57BL/6 mice, when B6.Gpi-1a mice were used as donors, was determined by analyzing the percentage of donor-derived hemoglobin or Gpi-1 in peripheral blood obtained from tail veins as previously described.25 To examine multilineage engraftment in these mice, whole-blood samples obtained from the retro-orbital sinus were stained separately with FITC-conjugated CD45R/B220, or FITC-conjugated CD3, and PE-conjugated Gr-1, lysed, and then isolated by flow cytometric cell sorting to yield B lymphocytes, T lymphocytes, and granulocytes, respectively. These purified lineages were then analyzed for Gpi-1 content.25Chimerism in C57BL/6 mice receiving donor cells of B6 BoyJ
origin was determined by means of flow cytometry to calculate the percentage of CD45.2 Short-term culture Cultures of 0.2 × 103 to 4 × 103 Sca-1+lin AM+ or AM
cells, or cell cycle subfractions of these cells, were initiated in
IMDM supplemented with 20% FBS and 2 × 105 mol/L
2-mercaptoethanol in an atmosphere of 5% CO2 in
100%-humidified air. Cytokines were delivered on day 0 as follows: 100 ng/mL mSCF (PeproTech), 500 U/mL mIL1 (Genzyme, Cambridge, MA), 100 U/mL mIL3 (Genzyme), 100 ng/mL hIL6, and 50 ng/mL human Flt3 ligand (hFlt3-L). The hIL6 and hFlt3-L were kind gifts from Amgen
(Thousand Oaks, CA) and Immunex (Seattle, WA), respectively. Care was
taken to keep the cell concentration below 1 × 106
cells/mL. Fresh cells or aliquots of cultured cells were removed on
days 1 and 2 and stained with propidium iodide for cell cycle analysis23 or incubated for an additional 11 to 14 days, at which time cells were harvested, enumerated, and analyzed by flow cytometry for the percentage of cells expressing Sca-1 with the use of
anti-Sca-1-PE.
Statistical analysis Data are expressed as the mean ± SEM where applicable. Differences between groups were analyzed by means of an unpaired 2-sided t test. A probability value of less than .05 was considered significant. Regression analysis was used to analyze the rate of exit from G0/G1 phases of the cell cycle.
Expression of AMs on Sca-1+lin cells. As seen in Table 1,
expression of these 6 AMs was varied, with nearly 100% of
Sca-1+lin cells expressing CD44 and CD49d,
and approximately 50% expressing CD49e and CD62L. Expression of CD11a
and CD43 was observed on approximately 80% of
Sca-1+lin cells.
Primitive and mature progenitor cell content of
Sca-1+lin AM+ or AM
cells was determined. Data in Table 2
show that CD43 and CD49e were expressed on the majority of HPP-CFCs
residing in the Sca-1+lin cell fraction,
while Sca-1+lin cells lacking expression of
either CD11a or CD62L contained a higher fraction of HPP-CFCs than
Sca-1+lin CD11a+ or
Sca-1+lin CD62L+ cells (Table 2).
LPP-CFCs, progenitors more committed to lineage differentiation than
HPP-CFCs, were mostly enriched among the same AM
profile as HPP-CFCs (Table 2). To ensure that the AM antibody used for cell sorting did not induce any negative
influences on primitive HPC clonogenic activity,
Sca-1+lin cells treated with each AM antibody
but not sorted on adhesion phenotype were also assayed for HPP- and
LPP-CFC content. HPP- and LPP-CFC activity of these antibody-treated
cells was not different from that of untreated
Sca-1+lin cells (data not shown), indicating
that the AM antibodies used for cell sorting did not inhibit in vitro
activity of progenitor cells. Fractions enriched for HPP- and LPP-CFC
activity also exhibited greater cellular expansion during 11 days of in
vitro cytokine-stimulated cell culture (Table 2).
Chimerism in mice transplanted with
Sca-1+lin AM+ or AM
cells, a competitive repopulation assay in lethally irradiated recipient mice was performed. Chimerism in recipient mice transplanted with either total Sca-1+lin cells or
Sca-1+lin AM+ or AM
cells was evident 4 weeks posttransplantation and continued to increase
until 2 months posttransplantation, at which time chimerism stabilized
(data not shown). Figure 2A shows that
Sca-1+lin cells expressing CD43 and CD49e
appeared to be highly enriched for long-term engraftment potential, as
Sca-1+lin cells lacking expression of either
of these 2 molecules failed to provide measurable chimerism in
recipients at 6 months posttransplantation. Sca-1+lin cells expressing low levels of
CD11a, CD49d, and CD62L were superior competitors compared with their
counterparts expressing higher levels (Figure 2A). Expression of CD44
did not appear to significantly correlate with enhanced or diminished
engraftment potential of Sca-1+lin cells. As
indicated by the horizontal bars in Figure 2A, anti-AM antibodies used
for cell sorting did not interfere with engraftment of
Sca-1+lin cells, as mice transplanted with
Sca-1+lin cells treated with each
anti-AM antibody exhibited chimerism similar to that of
mice receiving untreated Sca-1+lin cells. All
groups of Sca-1+lin
AM+ or AM cells contributed equally to
lineage-specific hematopoiesis, as indicated by similar levels of
donor-derived chimerism in myeloid (Gr-1+) and lymphoid
(CD3+ or B220+) cells (data not
shown).
The importance of CD49e expression in fractionating
Sca-1+lin Analysis of AM expression on engrafting vs nonengrafting phenotypes
of Sca-1+lin cells
demonstrated as having enriched engraftment potential were surveyed for
their repertoire of AM expression in comparison with their
nonengrafting counterparts. Figure 3
shows that expression of CD11a (panel A), CD49dbright
(panel D), and CD62L (panel F) was slightly lower on engrafting than on
nonengrafting phenotypes, while expression of CD43 and CD49e was
greater on engrafting cells. Interestingly, expression of CD43 (panel
B) was more than 4 times greater on engrafting phenotypes defined by
CD11a and CD49e, and expression of CD49e (Panel E) was more than 2- and
5-fold greater on engrafting phenotypes defined by CD11a and CD43,
respectively; this illustrates once again the importance of CD43 and
CD49e in engraftment. Examination of mean channel fluorescence and peak
channel fluorescence also revealed high levels of expression of CD43
and CD49e on engrafting phenotypes of
Sca-1±lin cells (data not shown). Expression
of CD44bright (Figure 3C) was slightly increased on
engrafting phenotypes.
Cell cycle status Since HSCs are believed to represent relatively quiescent cells, the cell cycle status of the 12 phenotypes of Sca-1+lin AM+ or AM
cells was evaluated to determine whether each of the AM subfractions of
Sca-1+lin cells enriched for engraftment
potential would also be more quiescent than the corresponding
nonengrafting phenotype. However, 5 out of 6 Sca-1+lin fractions that were negative or dim
for expression of AMs were significantly more quiescent than the
AM+ fraction (Figure 4). In
the cases of CD49d and CD62L, the engrafting phenotype was more
quiescent than the nonengrafting, but in the cases of CD43 and CD49e,
the engrafting phenotype contained the higher percentage of cycling
cells (Figure 4).
The cell cycle status of Sca-1+lin Cell cycle progression of AM subfractions of
Sca-1+lin AM+ or
AM cells (Figure 2). Differences in proliferative
potential between engrafting and nonengrafting phenotypes may be
related to the rate at which these cells exit
G0/G1 in vitro, as shown in Figure 5. AM fractions enriched for engraftment
potential, regardless of their relative degree of quiescence on day 0, exited G0/G1 more rapidly than nonengrafting
phenotypes (Figure 5). Cell cycle progression of
Sca-1+lin cells subfractionated with the use
of both CD49e and CD49d also correlated with the engraftment potential
of these cells. Sca-1+lin cells most enriched
for engraftment potential among these 4 phenotypes (CD49e+/CD49ddim) exited
G0/G1 more rapidly (slope =  25) than cells
lacking expression of CD49e (slopes = 2 to 11), or
Sca-1+lin cells expressing both CD49e and
CD49d cells (slope = 16).
Engraftment potential of Sca-1+lin cells (Figure 4) led us to question
whether quiescent or cycling cells within the AM-defined engrafting
phenotypes were contributing to engraftment potential and in vitro
hematopoiesis of these cells. To this end, engrafting phenotypes of
Sca-1+lin AM+ or AM
cells were subfractionated into G0/G1 and
S/G2+M fractions with the use of Hoechst 33342 and examined
for their engraftment potential. Figure 6
shows chimerism in mice transplanted with G0/G1
or S/G2 + M fractions of total
Sca-1+lin cells,
Sca-1+lin CD11a,
Sca-1+lin CD49ddim, and
Sca-1+lin CD49e+ cells.
G0/G1 cells, regardless of which AM was used in
fractionation, provided greater levels of chimerism than equal numbers
of S/G2+M cells (P < .005), illustrating the
enhanced engraftment potential of quiescent cells.
G0/G1 cells also provided higher levels of chimerism than equal numbers of total
Sca-1+lin AM subfraction (data not shown),
negating any suggestion that the Hoechst dye may negatively influence
the function of primitive HPCs in active phases of cell cycle. Of
interest in these analyses is that, although a relatively large
percentage of Sca-1+lin CD49e+
cells were in active phases of cell cycle (Figure 4), only those in
G0/G1 accounted for the majority of the
engraftment potential of this phenotype. All groups of cells
contributed equally to lineage-specific hematopoiesis (myeloid and
lymphoid; data not shown). Table 3 shows
that the quiescent fractions of engrafting Sca-1+lin cells possessed greater
proliferative potential and gave rise to greater numbers of
Sca-1+ cells during 14 days of in vitro cytokine-stimulated
cell culture than engrafting cells in active phases of cell
cycle.
In this report, we define a theoretical phenotype of a
Sca-1+lin To the best of our knowledge, antibodies used in these studies were
nonblocking. In mice where long-term engraftment potential was
predominantly among AM Although much work has been performed illustrating the importance of
several AMs in various aspects of stem cell function, AMs involved in
the sequential movement of transplanted or mobilized HSCs through BM
endothelium to or from the periphery are not yet defined. It is likely
that different AMs are involved at different stages of stem cell
homing, from initial tethering of HSCs on BM endothelium, to rolling,
attachment, passage through endothelial barriers, movement within BM
tissues, and final anchorage within specialized BM niches, similar to
that described for lymphocyte trafficking and leukocyte
emigration.31 Mobilization could conceivably involve
similar AMs but in reverse order. We found that a sizable fraction of
Sca-1+lin While a role of CD49d in homing to and egress from the BM of
clonogenic progenitors28,37,38 and malignant
cells39,40 has been well defined, to date no one has
directly examined CD49d in homing of long-term engrafting cells. The
laboratory of Dr Papayannopoulou has shown the ability of
function-blocking anti-CD49d antibody to mobilize murine long-term
engrafting cells7 and, more recently, to possess a role in
homing and mobilization of human CD34+ cells in fetal
sheep.1 Given the complexity of homing, it is possible
that CD49d may not be involved in the initial steps of HSC homing, but
may be critical for the lodgment of HSCs within BM microenvironment
following homing via different AMs. This notion is supported by our
findings demonstrating that Sca-1+lin CD62L expression on human mobilized peripheral blood (MPB) CD34+ cells has been shown to correlate with rapid platelet engraftment,4,44 but was found in our studies to be detrimental to engraftment potential, presenting the possibility that short-term engrafting cells may express CD62L but long-term engrafting cells may not. Such a concept, however, could be in direct conflict with that recently presented by Ziljmans et al,45 who found primitive HPCs to be responsible for both short- and long-term engraftment. Additionally, since the AM profile is reportedly different in human BM and MPB CD34+ cells,4,5,44,46-49 it should not be surprising to find differences in the AM profiles of long-term reconstituting cells from either BM or MPB, within or outside species barriers. A surprising finding in our studies was the rapid in vitro proliferation and cycling of long-term engrafting cells, which are generally believed to be quiescent.8 While it is possible that cytokine-responsive cells in vitro were not the same cells contributing to long-term engraftment, several lines of evidence support the notion that murine long-term engrafting cells cycle rapidly, at least following transplantation.15,27,50 The rate of in vivo proliferation of colony-forming unit-spleen has been demonstrated to predict long-term engraftment potential of transplanted HPC.27,50 In addition, Nilsson et al15 demonstrated cycling of engrafting murine HSCs within the first 12 hours posttransplantation. Collectively, these data suggest that rapid proliferative responses to cytokine stimulation, either in vivo or in vitro, may be predictive of engraftment potential of selected groups of murine primitive HPCs. The relationship between cell cycle position and engraftment
capability of HSCs has been the focus of much
investigation.12,13,15,51 Sca-1+lin
Submitted November 5, 1999; accepted April 14, 2000.
Supported by National Institutes of Health grants RO1 HL55716 and RO1 HL62200 (to E.F.S.); Herman B Wells Center for Pediatric Research is a Center of Excellence in Molecular Hematology (NIDDK P30 DK49218).
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: Christie M. Orschell-Traycoff, Indiana University School of Medicine, 1044 West Walnut St, R4-202, Indianapolis, IN 46202-5254; e-mail: ctraycof{at}iupui.edu.
1.
Zanjani E, Flake A, Almeida-Porada G, Tran N, Papayannopoulou T.
Homing of human cells in the fetal sheep model: modulation by antibodies activating or inhibiting very late activation antigen-4-dependent function.
Blood.
1999;94:2515 2. Liesveld JL, Dipersio JF, Abboud CN. Integrins and adhesive receptors in normal and leukemic CD34+ progenitor cells: potential regulatory checkpoints for cellular traffic. Leuk Lymphoma. 1994;14:19[Medline] [Order article via Infotrieve]. 3. van der Loo JCM, Xiao X, McMillin D, Hashino K, Kato I, Williams DA. VLA-5 is expressed by mouse and human long-term repopulating hematopoietic cells and mediates adhesion to extracellular matrix protein fibronectin. J Clin Invest. 1998;102:1051[Medline] [Order article via Infotrieve].
4.
Dercksen MW, Gerritsen WR, Rodenhuis S, et al.
Expression of adhesion molecules on CD34+ cells: CD34+ L-selectin+ cells predict a rapid platelet recovery after peripheral blood stem cell transplantation.
Blood.
1995;85:3313 5. Leavesley DI, Oliver JM, Swart BW, Berndt MC, Haylock DN, Simmons PJ. Signals from platelet/endothelial cell adhesion molecule enhance the adhesive activity of the very late antigen-4 integrin of human CD34+ hemopoietic progenitor cells. J Immunol. 1994;153:4673[Abstract].
6.
Lewinsohn D, Nagler A, Ginzton N, Greenberg P, Butcher E.
Hematopoietic progenitor cell expression of the H-CAM (CD44) homing-associated adhesion molecule.
Blood.
1990;75:589
7.
Craddock C, Nakamoto B, Andrews R, Priestley G, Papayannopoulou T.
Antibodies to VLA4 integrin mobilize long-term repopulating cells and augment cytokine-induced mobilization in primates and mice.
Blood.
1997;90:4779
8.
Ogawa M.
Differentiation and proliferation of hematopoietic stem cells.
Blood.
1993;81:2844 9. Bradford G, Williams B, Rossi R, Bertoncello I. Quiescence, cycling, and turnover in the primitive hematopoietic stem cell compartment. Exp Hematol. 1997;25:445[Medline] [Order article via Infotrieve]. 10. Abkowitz JL, Catlin SN, Guttorp P. Evidence that hematopoiesis may be a stochastic process in vivo. Nat Med. 1996;2:190[Medline] [Order article via Infotrieve].
11.
Shah AJ, Smogorzewska EM, Hannum C, Crooks GM.
Flt3 ligand induces proliferation of quiescent human bone marrow CD34+CD38
12.
Gothot A, van der Loo J, Clapp W, Srour E.
Cell cycle-related changes in repopulating capacity of human mobilized peripheral blood CD34+ cells in non-obese diabetic/severe combined immune-deficient mice.
Blood.
1998;92:2641 13. Traycoff CM, Cornetta K, Yoder MC, Davidson A, Srour EF. Ex vivo expansion of murine hematopoietic progenitor cells generates classes of expanded cells possessing varying levels of bone marrow repopulating potentials. Exp Hematol. 1996;24:299[Medline] [Order article via Infotrieve].
14.
Peters SO, Kittler ELW, Ramshaw HS, Quesenberry PJ.
Ex vivo expansion of murine marrow cells with interleukin-3 (IL-3), IL-6, IL-11, and stem cell factor leads to impaired engraftment in irradiated hosts.
Blood.
1996;87:30
15.
Nilsson SK, Dooner MS, Quesenberry PJ.
Synchronized cell-cycle induction of engrafting long-term repopulating stem cells.
Blood.
1997;90:4646 16. Becker PS, Nilsson SK, Li Z, et al. Adhesion receptor expression by hematopoietic cell lines and murine progenitors: modulation by cytokines and cell cycle status. Exp Hematol. 1999;27:533[Medline] [Order article via Infotrieve].
17.
Yamaguchi M, Ikebuchi K, Hirayama F, et al.
Different adhesive characteristics and VLA-4 expression of CD34+ progenitors in G0/G1 versus S+G2/M phases of the cell cycle.
Blood.
1998;92:842 18. Traycoff C, Yoder M, Hiatt K, Srour E. Cell cycle stage-specific expression of adhesion molecules may augment engraftment potential of quiescent but not mitotically active hematopoietic progenitor cells [abstract]. Blood. 1996;88:475. 19. Srour E, Traycoff C. Mobilization of primitive hematopoietic progenitor cells into the periphery may involve their exit from the bone marrow microenvironmnet without entry into active phases of the cell cycle [abstract]. Blood. 1996;88:532. 20. Traycoff CM, Hoffman R, Zanjani ED, et al. Measurement of marrow repopulating potential of human hematopoietic progenitor and stem cells using a fetal sheep model. Prog Clin Biol Res. 1994;389:281[Medline] [Order article via Infotrieve]. 21. Leemhuis T, Yoder MC, Grigsby S, Aguero B, Eder P, Srour E. Isolation of primitive human bone marrow hematopoietic progenitor cells using Hoechst 33342 and Rhodamine 123. Exp Hematol. 1996;24:1215[Medline] [Order article via Infotrieve].
22.
Uchida N, Friera A, He D, Reitsma M, Tsukamoto A, Weissman I.
Hydroxyurea can be used to increase mouse c-kit+, Thy-1.1lo, Lin
23.
Srour EF, Brandt JE, Leemhuis T, Ballas CB, Hoffman R.
Relationship between cytokine-dependent cell cycle progression and MHC class II antigen expression by human CD34+ HLA-DR
24.
Yoder MC, King B, Hiatt K, Williams DA.
Murine embryonic yolk sac cells promote in vitro proliferation of bone marrow high proliferative potential colony-forming cells.
Blood.
1995;86:1322 25. Harrison D, Jordan C, Zhong R, Astle C. Primitive hemopoietic stem cells: direct assay of most productive populations by competitve repopulation with simple binomial correlation and covariance calculations. Exp Hematol. 1993;21:206[Medline] [Order article via Infotrieve]. 26. Wolf NS, Kone A, Priestley GV, Bartelmez SH. In vivo and in vitro characterization of long-term repopulating primitive hematopoietic cells isolated by sequential Hoechst 33342-rhodamine 123 FACS selection. Exp Hematol. 1993;21:614[Medline] [Order article via Infotrieve].
27.
Neben S, Marcus K, Mauch P.
Mobilization of hematopoietic stem and progenitor cell subpopulations from the marrow to the blood of mice following cyclophosphamide and/or granulocyte colony-stimulating factor.
Blood.
1993;81:1960
28.
Vermeulen M, Le Pesteur F, Gagnerault M, Mary J, Sainteny F, Lepault F.
Role of adhesion molecules in the homing and mobilization of murine hematopoietic stem and progenitor cells.
Blood.
1998;92:894 29. Gallatin W, Weissman I, Butcher E. A cell surface molecule involved in organ-specific homing of lymphocytes. Nature. 1983;304:30[Medline] [Order article via Infotrieve].
30.
Carlos T, Harlan J.
Leukocyte-endothelial adhesion molecules.
Blood.
1994;84:2068 31. Springer T. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994;76:301[Medline] [Order article via Infotrieve]. 32. Craddock C, Nakamoto B, Elices M, Papayannapoulou T. The role of CS1 moiety of fibronectin in VLA4-mediated haemopoietic progenitor trafficking. Br J Haematol. 1997;97:15[Medline] [Order article via Infotrieve].
33.
Bazil V, Brandt J, Chen S, et al.
A monoclonal antibody recognizing CD43 (leukosialin) initiates apoptosis of human hematopoietic progenitor cells but not stem cells.
Blood.
1996;87:1272 34. Hirayama F, Ogawa M. CD43 expression by murine lymphohemopoietic progenitors. Int J Hematol. 1994;60:191[Medline] [Order article via Infotrieve]. 35. Moore T, Huang S, Terstappen LW, Bennett M, Kumar V. Expression of CD43 on murine and human pluripotent hematopoietic stem cells. J Immunol. 1994;153:4978[Abstract].
36.
Verfaillie CM, Benis A, Iida J, McGlave PB, McCarthy JB.
Adhesion of committed human hematopoietic progenitors to synthetic peptides from the C-terminal heparin-binding domain of fibronectin: cooperation between the integrin alpha 4 beta 1 and the CD44 adhesion receptor.
Blood.
1994;84:1802
37.
Papayannopoulou T, Nakamoto B.
Peripheralization of hemopoietic progenitors in primates treated with anti-VLA4 integrin.
Proc Natl Acad Sci U S A.
1993;90:9374
38.
Papayannopoulou T, Craddock C, Nakamoto B, Priestley G, Wolf N.
The VLA4/VCAM adhesion pathway defines contrasting mechanisms of lodgement of transplanted murine hemopoietic progenitors between bone marrow and spleen.
Proc Nat Acad Sci U S A.
1995;92:9647 39. Filshie R, Gottlieb D, Bradstock K. VLA-4 is involved in the engraftment of the human pre-B acute lymphoblastic leukemia cell line NALM-6 in SCID mice. Br J Haematol. 1998;102:1292[Medline] [Order article via Infotrieve]. 40. Matsuura N, Purzon-McLaughlin W, Irie A, Morikawa Y, Kakudo K, Takada Y. Induction of experimental bone metastasis in mice by transfection of integrin alpha 4 beta 1 into tumor cells. Am J Pathol. 1996;148:55[Abstract]. 41. Prosper F, Stroncek D, McCarthy JB, Verfaillie CM. Mobilization and homing of peripheral blood progenitors is related to reversible downregulation of alpha4 beta1 integrin expression and function. J Clin Invest. 1998;101:2456[Medline] [Order article via Infotrieve].
42.
Kovach NL, Lin N, Yednock T, Harlan JM, Broudy VC.
Stem cell factor modulates avidity of alpha 4 beta 1 and alpha 5 beta 1 integrins expressed on hematopoietic cell lines.
Blood.
1995;85:159
43.
Levesque JP, Leavesley DI, Niutta S, Vadas M, Simmons PJ.
Cytokines increase human hemopoietic cell adhesiveness by activation of very late antigen (VLA)-4 and VLA-5 integrins.
J Exp Med.
1995;181:1805 44. Watanabe T, Dave B, Heimann D, Jackson J, Kessinger A, Talmadge J. Cell adhesion molecule expression on CD34+ cells in grafts and time to myeloid and platelet recovery after autologous stem cell transplantation. Exp Hematol. 1998;26:10[Medline] [Order article via Infotrieve].
45.
Ziljmans J, Visser J, Laterveer L, et al.
The early phase of engraftment after murine blood cell transplantation is mediated by hematopoietic stem cells.
Proc Nat Acad Sci U S A.
1998;95:725 46. Turner ML, McIlwaine K, Anthony RS, Parker AC. Differential expression of cell adhesion molecules by human hematopoietic progenitor cells from bone marrow and mobilized adult peripheral blood. Stem Cells. 1995;13:311[Abstract].
47.
To LB, Haylock DN, Dowse T, et al.
A comparative study of the phenotype and proliferative capacity of peripheral blood (PB) CD34+ cells mobilized by four different protocols and those of steady-phase PB and bone marrow CD34+ cells.
Blood.
1994;84:2930 48. Fultz C, Shivers S, Smilee R, Janssen W. Hematopoietic cell adhesion molecule and very late antigen-4 but not L-selectin are differentially expressed between marrow and blood [abstract]. Exp Hematol. 1996;24:1078. 49. Zini J, Marolleau J, Miclea J, Mouton V, Benbunan M. Expression of adhesion molecules on CD34+ cells: comparative studies between peripheral blood and bone marrow stem cells [abstract]. Exp Hematol. 1996;24:1034.
50.
Mauch P, Hellman S.
Loss of hematopoietic stem cell self-renewal after bone marrow transplantation.
Blood.
1989;74:872
51.
Fleming WH, Alpern EJ, Uchida N, Ikuta K, Spangrude GJ, Weissman IL.
Functional heterogeneity is associated with the cell cycle status of murine hematopoietic stem cells.
J Cell Biol.
1993;122:897
52.
Sato T, Laver J, Ogawa M.
Reversible expression of CD34 by murine hematopoietic stem cells.
Blood.
1999;94:2548
53.
Orlic D, Fischer R, Nishikawa S, Nienhuis AW, Bodine DM.
Purification and characterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor.
Blood.
1993;82:762
© 2000 by The American Society of Hematology.
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  D. Daria, M.-D. Filippi, E. S. Knudsen, R. Faccio, Z. Li, T. Kalfa, and H. Geiger The retinoblastoma tumor suppressor is a critical intrinsic regulator for hematopoietic stem and progenitor cells under stress Blood, February 15, 2008; 111(4): 1894 - 1902. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
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|
|
|
|
|
|
Y. Si, S. Ciccone, F.-C. Yang, J. Yuan, D. Zeng, S. Chen, H. J. van de Vrugt, J. Critser, F. Arwert, L. S. Haneline, et al. Continuous in vivo infusion of interferon-gamma (IFN-{gamma}) enhances engraftment of syngeneic wild-type cells in Fanca-/- and Fancg-/- mice Blood, December 15, 2006; 108(13): 4283 - 4287. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Nygren, D. Bryder, and S. E. W. Jacobsen Prolonged Cell Cycle Transit Is a Defining and Developmentally Conserved Hemopoietic Stem Cell Property J. Immunol., July 1, 2006; 177(1): 201 - 208. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
R. H. Lee, S. C. Hsu, J. Munoz, J. S. Jung, N. R. Lee, R. Pochampally, and D. J. Prockop A subset of human rapidly self-renewing marrow stromal cells preferentially engraft in mice Blood, March 1, 2006; 107(5): 2153 - 2161. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
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|
|
|
|
 |
|
 M. Rosu-Myles, E. Stewart, J. Trowbridge, C. Y. Ito, P. Zandstra, and M. Bhatia A unique population of bone marrow cells migrates to skeletal muscle via hepatocyte growth factor/c-met axis J. Cell Sci., October 1, 2005; 118(19): 4343 - 4352. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Liang, G. Van Zant, and S. J. Szilvassy Effects of aging on the homing and engraftment of murine hematopoietic stem and progenitor cells Blood, August 15, 2005; 106(4): 1479 - 1487. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. C. Zhang and H. F. Lodish Murine hematopoietic stem cells change their surface phenotype during ex vivo expansion Blood, June 1, 2005; 105(11): 4314 - 4320. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. E. Broxmeyer, C. M. Orschell, D. W. Clapp, G. Hangoc, S. Cooper, P. A. Plett, W. C. Liles, X. Li, B. Graham-Evans, T. B. Campbell, et al. Rapid mobilization of murine and human hematopoietic stem and progenitor cells with AMD3100, a CXCR4 antagonist J. Exp. Med., April 18, 2005; 201(8): 1307 - 1318. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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|
|
|
|
|
|
P. M. Chilton, F. Rezzoug, M. Z. Ratajczak, I. Fugier-Vivier, J. Ratajczak, M. Kucia, Y. Huang, M. K. Tanner, and S. T. Ildstad Hematopoietic stem cells from NOD mice exhibit autonomous behavior and a competitive advantage in allogeneic recipients Blood, March 1, 2005; 105(5): 2189 - 2197. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Chavakis, A. Aicher, C. Heeschen, K.-i. Sasaki, R. Kaiser, N. El Makhfi, C. Urbich, T. Peters, K. Scharffetter-Kochanek, A. M. Zeiher, et al. Role of {beta}2-integrins for homing and neovascularization capacity of endothelial progenitor cells J. Exp. Med., January 3, 2005; 201(1): 63 - 72. [Abstract] [Full Text] [PDF]
|
|
|
|
|
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Y. Huang, F. Rezzoug, P. M. Chilton, H. L. Grimes, D. E. Cramer, and S. T. Ildstad Matching at the MHC class I K locus is essential for long-term engraftment of purified hematopoietic stem cells: a role for host NK cells in regulating HSC engraftment Blood, August 1, 2004; 104(3): 873 - 880. [Abstract] [Full Text] [PDF]
|
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S. S. Perry, H. Wang, L. J. Pierce, A. M. Yang, S. Tsai, and G. J. Spangrude L-selectin defines a bone marrow analog to the thymic early T-lineage progenitor Blood, April 15, 2004; 103(8): 2990 - 2996. [Abstract] [Full Text] [PDF]
|
|
|
|
|
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F. Ahmed, S. J. Ings, A. R. Pizzey, M. P. Blundell, A. J. Thrasher, H. T. Ye, A. Fahey, D. C. Linch, and K. L. Yong Impaired bone marrow homing of cytokine-activated CD34+ cells in the NOD/SCID model Blood, March 15, 2004; 103(6): 2079 - 2087. [Abstract] [Full Text] [PDF]
|
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P. A. Plett, S. M. Frankovitz, and C. M. Orschell Distribution of marrow repopulating cells between bone marrow and spleen early after transplantation Blood, September 15, 2003; 102(6): 2285 - 2291. [Abstract] [Full Text] [PDF]
|
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J.-F. Lambert, M. Liu, G. A. Colvin, M. Dooner, C. I. McAuliffe, P. S. Becker, B. G. Forget, S. M. Weissman, and P. J. Quesenberry Marrow Stem Cells Shift Gene Expression and Engraftment Phenotype with Cell Cycle Transit J. Exp. Med., June 2, 2003; 197(11): 1563 - 1572. [Abstract] [Full Text] [PDF]
|
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|
|
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P. Denning-Kendall, S. Singha, B. Bradley, and J. Hows Cytokine Expansion Culture of Cord Blood CD34+ Cells Induces Marked and Sustained Changes in Adhesion Receptor and CXCR4 Expressions Stem Cells, January 1, 2003; 21(1): 61 - 70. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. J. Quesenberry, G. A. Colvin, and J.-F. Lambert The chiaroscuro stem cell: a unified stem cell theory Blood, December 15, 2002; 100(13): 4266 - 4271. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. A. Plett, S. M. Frankovitz, and C. M. Orschell-Traycoff In vivo trafficking, cell cycle activity, and engraftment potential of phenotypically defined primitive hematopoietic cells after transplantation into irradiated or nonirradiated recipients Blood, November 15, 2002; 100(10): 3545 - 3552. [Abstract] [Full Text] [PDF]
|
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|
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N. Askenasy and D. L. Farkas Optical Imaging of PKH-Labeled Hematopoietic Cells in Recipient Bone Marrow In Vivo Stem Cells, November 1, 2002; 20(6): 501 - 513. [Abstract] [Full Text] [PDF]
|
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|
C. E. Muller-Sieburg, R. H. Cho, M. Thoman, B. Adkins, and H. B. Sieburg Deterministic regulation of hematopoietic stem cell self-renewal and differentiation Blood, July 30, 2002; 100(4): 1302 - 1309. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Giet, D. R. Van Bockstaele, I. Di Stefano, S. Huygen, R. Greimers, Y. Beguin, and A. Gothot Increased binding and defective migration across fibronectin of cycling hematopoietic progenitor cells Blood, March 15, 2002; 99(6): 2023 - 2031. [Abstract] [Full Text] [PDF]
|
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|
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A. Jetmore, P. A. Plett, X. Tong, F. M. Wolber, R. Breese, R. Abonour, C. M. Orschell-Traycoff, and E. F. Srour Homing efficiency, cell cycle kinetics, and survival of quiescent and cycling human CD34+ cells transplanted into conditioned NOD/SCID recipients Blood, March 1, 2002; 99(5): 1585 - 1593. [Abstract] [Full Text] [PDF]
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Y. J. Summers, C. M. Heyworth, E. A. de Wynter, J. Chang, and N. G. Testa Cord Blood G0 CD34+ Cells Have A Thousand-Fold Higher Capacity For Generating Progenitors In Vitro Than G1 CD34+ Cells Stem Cells, November 1, 2001; 19(6): 505 - 513. [Abstract] [Full Text]
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S. J. Szilvassy, T. E. Meyerrose, P. L. Ragland, and B. Grimes Differential homing and engraftment properties of hematopoietic progenitor cells from murine bone marrow, mobilized peripheral blood, and fetal liver Blood, October 1, 2001; 98(7): 2108 - 2115. [Abstract] [Full Text] [PDF]
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| Copyright © 2000 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
cells fractionated on the basis of
adhesion molecule expression and position in cell cycle
Top
Abstract
Introduction
),
CD43 (
), CD49d (
), CD49e (
), and CD62L (
) to give
engrafting or nonengrafting phenotypes, designated E and NE,
respectively. CD44 was omitted in the primary sort since this marker
was not useful in defining engrafting phenotypes (Figure 
), or engrafting
phenotypes (dotted lines) and nonengrafting phenotypes (solid lines) of
Sca-1+lin