1. Art and Diseño

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RAPID COMMUNICATION
~~
~
~~~~~
Peripheral Blood Progenitor Cells Mobilized by Recombinant Human
Granulocyte Colony-Stimulating Factor Plus Recombinant Rat Stem Cell
Factor Contain Long-Term Engrafting Cells Capable
of Cellular Proliferation for More Than Two Years as Shown
by Serial Transplantation in Mice
By X.-Q. Yan, C. Hartley, P. McElroy, A. Chang, C. McCrea, and I. McNiece
Mobilized peripheral bloodprogenitor cells (PBPC) have
been shown t o provide rapidengraftment in patients given
high-dose chemotherapy. PBPC contain cells with long-term
engraftment potential as shown in animal models. In this
study we have further analyzed mobilized PBPC for their
ability t o support serial transplantation of irradiated mice.
Transplantation of recombinant human granulocyte colonystimulating factor (rhG-CSF) plus recombinant rat stemcell
factor (rrSCF) mobilized PBPC resulted in 989’0 donor engraftment of primary recipients at 12 t o 14 months posttransplantation. Bone marrow (BM) cells from these primary
recipients were harvested and transplanted into secondary
recipients. At 6 months posttransplantation, all surviving
secondary recipients had donor engraftment. Polymerase
chain reaction(PCR) analysis showed greater than 909’0male
cells in spleens, thymuses, and lymphnodes. Myeloid colonies from BM cells of secondary recipients demonstrated
granulocyte/macrophage colony-forming cells (GM-CFC) of
male origin in all animals. In comparison, transplantation of
rhG-CSF mobilized PBPC resulted in decreased male engraftment in secondary recipients. BM cells from secondary
recipients, who originally received PBPC mobilized by the
combination ofrrSCF and rhG-CSF, were furtherpassagedt o
tertiary female recipients. At 6 months posttransplantation,
909’0 of animals had male-derived hematopoiesis by wholeblood PCR analysis. These data showed thatPBPC mobilized
with rhG-CSF plus rrSCF contained cells that are transplantable and able t o maintain hematopoiesis for more than 26
months, suggesting that the mobilized long-termreconstituting stemcells (LTRC) have extensive proliferative potential and resemble those that reside in the BM. In addition.
the data demonstrated increased mobilization of LTRC with
rhG-CSF plus rrSCF compared t o rhG-CSF alone.
0 1995 by The American Societyof Hematology.
H
were transplanted into lethally irradiated female mice. The
primary recipients had greater than 90%donor engraftment
at 12 to 14 months posttransplantation.’ BM cells from these
animals have been transplanted into secondary and tertiary
recipients and the male-derived hematopoiesis is evaluated.
IGH-DOSE CHEMOTHERAPY for treatment of solid
tumors requires cellular support of bone marrow (BM)
cells or peripheral blood progenitor cells (PBPC). BM transplantation (BMT) has been shown to provide long-term durable engraftment of recipients. PBPC transplantation has
primarily been performed in the autologous setting, complicating the determination of long-term engraftment of donor
cells from endogenous recovery. Mouse models have been
used to evaluate the long-term engraftment potential of
growth factor mobilized PBPC. Granulocyte colony-stimulating factor (rhG-CSF),’ stem cell factor (SCF),’ and interleukin-l (IL-l)3mobilized PBPC have been shown to provide donor engraftment in irradiated mice for 6 to 12 months
posttransplantation. In addition, we have shown that PBPC
mobilized by the combination of an optimal dose of rhGCSF plus low doses of recombinant rat SCF have increased
numbers of cells with short-term and long-term engraftment
potential compared with PBPC mobilized with rhG-CSF
al~ne.~,~
A number of studies have demonstrated that serial transplantation of BM cells leads to decreased long-term reconstitution ability and that BM cells from previously transplanted
animals have decreased long-term reconstituting ability
(LTRA) compared with BM cells from untreated anim a l ~ . Studies
~ . ~ by Harrison et a1 demonstrated that the decline in LTRA was not a function of age of the stem cell^.^^*
Moreover, they concluded that the proliferative potential of
stem cells was decreased after transplantation, probably because of differentiative pressure on the stem cells. In this
study we have evaluated the short-term and long-term reconstituting capacity ofBM cells from mice that were transplanted with PBPC mobilized with rhG-CSF alone or the
combination of rhG-CSF plus rrSCF. Male donor mice were
mobilized for 7 days with growth factors and the PBPC
Blood, Vol 85, No 9 (May l ) , 1995: pp 2303-2307
MATERIALS AND METHODS
Mice. Eight- to 12-week-old female and male splenectomized
(C57BW6J x DBN2J) F,(BDF,) mice were purchased from Charles
River Laboratories (Wilmington, MA). All animals were housed at
animal facilities of Amgen Inc (Thousand Oaks, CA) under sterile
conditions and were supplied with acidified water.
Transplantation. Mobilization of PBPC using BDF, mice has
been described previously! Briefly, male splenectomized BDF, mice
were treated with rhG-CSF at 200 pgkgld, or rhG-CSF 200 pg/kgl
d plus recombinant rat pegylated SCF (rrSCF-PEG) at 25 pg/kg/d
for 7 days. Both rhG-CSF and rrSCF-PEG were produced by Amgen
Inc.’.’’ BM cells from femurs and tibias were flushed in 2% fetal
bovine serum-Hanks’ balanced saline solution (FBSHBSS), washed,
and resuspended in isotonic saline containing 1 % bovine serum albumin (BSA). Nucleated cells were counted and transplanted into yirradiated (GI3’, 12 Gyat split dose 4 hours apart) female BDFl
mice.
From the Department of Developmental Hematology, Amgen Inc,
Thousand Oaks, CA.
Submitted December 21, 1994; accepted February 8, 1995.
Address reprint requests to I. McNiece, PhD, Amgen Inc. Amgen
Center, T-I-A-207, 1840 DeHavilland Dr, Thousand Oaks, CA
91320-1 789.
The publication costsof this article weredefrayed in part by page
charge payment. This article must therefore be hereby marked
“advertisement” in accordance with 18 U.S.C. section 1734 solely to
indicate this fact.
0 1995 by The American Society of Hematology.
0004-4971/95/8509-0043$3.00/0
2303
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YAN ET AL
2304
Table 1. Primary Recipients Transplanted With PBPC Mobilized
With rhG-CSF or rhG-CSF Plus nSCF
l
plus SCF 25 pgkgday
Injected
PBPC (2.5 x 105 LD Cells)
+
A (5)
B (10)
+
+ (I.slr*onyPCRI
1. Whole blood PCR
6 months
No. of Cells
Factors (sglkgld)
X
rhG-CSF (200)
52-59
rhG-CSF (200)
+ rrSCF (25)
105
2.5
2.5
96 Male Positive
Time
lwks)
Cells in
Whole Blood
48
>go%
>90%
Mice reconstituted with PBPC mobilized with either rhG-CSF alone
or rhG-CSF plus rrSCF were killed at the times indicated. Results of
donor (malekderived hematopoiesis are shown byPCR analysis of
whole blood and single colonies picked from BM cultures.
BM (0.1-1.0 femur)
+
Donor
Groups
(n)
3.SDI& Ivm & thv PCR
intron sequence.I4Amplified materials were electrophoresed in 1.5%
agarose, Southern blotted, and hybridized to 32P-end-labeledSry or
PDGF B receptor internal sequences (25-bp oligos), respectively,
with conventional hybridization protocols.I2 An overview of the serial transplantation and analysis is presented in Fig 1.
BM (0.1-1 .O femur)
+
RESULTS
PBPC mobilized with rhG-CSF alone (group A) or rhGCSF plus rrSCF (group B) were transplanted into lethally
2xgoorads
irradiated mice at 2.5 X IO' low-density cells per animal.
6 months
At 48-59 weeks posttransplantation, these mice had greater
than 90% donor (male) cells in the peripheral blood (Table
I Whole blood PCR 1
1) and 81% (group A) and 98% (group B) donor-derived
GM-CFC in the BM.* The BM cells from these animals
Fig 1. Experimental protocol showing serial transplantation of
were harvested as described in Materials and Methods and
PBPC mobilized with rhG-CSF alone (A) or rhG-CSF plus rrSCF (B) to
transplanted into secondary female recipients, and these aniprimary, secondary and tertiary recipients.
mals were analyzed at 26 weeks posttransplantation (Table
2). No difference was observed in the BM cellularities of
secondary recipient mice in both groups of animals (data not
Colony assays. Nucleated BM cells, 2 X 104, were plated in 1.0
shown). Mice from groups 1 and 2 had 67% and 73% surmL Iscove's modified Dulbecco's medium (IMDM) plus methyl
cellulose containing 10%FBS, I % BSA, lipids, transferrin, in~u1in.l~ vival at 40 weeks, respectively, whereas mice in groups 3
2.5 ng/mL rmlL-3, 2.5 ng/mL rhlL-10, 1 0 0 ng/mL rrSCF, 1 0 0 ng/
and 4 had 87% and 97% survival, respectively (Table 2).
mL rhlL-6, 1 0 0 ng/mL rhIL-11, and 2 UlmL rh erytropoietin
Whole-blood PCR analysis of the long-term surviving mice
(rhEpo). Granulocytelmacrophage (GM) and multilineage colonies
showed 100% of groups 3 and 4 mice were positive for
were scored after 9 days of incubation in 5% CO2 at 37°C.
the Y chromosome and 40% and 91% of groups 1 and 2,
Y-chromosomepolymerase chain reaction (PCR) analysis. PCR
respectively. PCR analysis was performed on individual colanalysis of male-derived hematopoiesis in whole blood and single
onies picked from BM cultures to assess male-derived hemacolonies has been described in detail previously.2 Semi-quantitative
topoietic
precursors in the secondary recipients. This apPCR was performed on spleens, thymuses, and lymph nodes to
proach is a better estimate of donor-derived hematopoiesis
measure the percentage of male-derived cells. Briefly, cell suspenthan whole-bloodPCR
analysis because of variables in
sions were prepared by breaking spleens, thymuses, and lymphoid
-9
+
nodes in a 100-pm cell strainer (Falcon, Lincoln Park, NJ) with a
plunger from a 1.0-mL syringe in 2% FBS HBSS. Cells were than
lysed anddigested with proteinase K. DNA was extracted by conventional phenol/chloroform methods.I2 One hundred nanograms of
DNA was amplified in 200 pL thin-wall tubes (Perkin Elmer Cetus,
Norwalk, C T ) in a total volume of 50 pL reaction buffer containing
10 pmol of mouse Y-chromosome-specific primers, or mouse platelet-derived growth factor (PDGF) B receptor specific primers, 1.5
mmol/L MgCI2. 50 mmol/L KCI, 10 mmol/L Tris-HCI, pH 8.3,0.2
mmol/L dNTPs, and 2.5 U Taq polymerase (Boehringer Mannheim,
Indianapolis, IN). PCR reaction was initiated at 94°C for 4 minutes,
followed by 28 cycles at 94°C for 1 minute, 62°C for 1 minute, and
72°C for 2 minutes. Amplification with Y-specific primers resulted
in a 722-bp fragment corresponding to the Sry locus sequence 256978," whereas amplification with PDGF B receptor primers generatedan approximately 750-bp fragment corresponding to mouse
PDGF B receptor cDNA sequence 948-1166,which contains an
Table 2. Transplantation of BM Cells From Primary Recipient Mice
Into Secondary Recipient Mice
Recipient
Injected
Groups
(donor)
1. (A)
2.(A)
3.(B)
4.
(B)
Cells
No. of
x
106
1.o
8.4 2 1.8
1.o
9.2 2 1.7
Long-Term
Time PostSurvivOrSl
Positive1
transplantation
Mice
lwks)
Survivors
Transplanted
40
35
26/30
(0.87)
26
29/30
(0.97)
26
10/15 (0.67)
11/15 (0.73)
No. of Male
Long-Term
4/10 (0.40)
10/11 (0.91)
26/26(1.0)
29/29(1.0)
Mice in groups 1 and 2 received BM cells from donor group A. Mice
in groups 3 and 4 received cells from donor group B. Mice were
analyzed at 26 to 40 weeks posttransplantation. Results show PCR
analysis of long-term surviving animals.
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MOBILIZATION OF LONG-TERM ENGRAFTINGCELLS
100
=
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1
2305
75
0
0
.m
g
a
0
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0
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H
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ccc
25
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s
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03
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a
Gp4
Gp3
Gp2
Gpl
Fig 2. Detection of donor-derivedhematopoiesisin secondary recipient mice. Secondary recipients were killed at 26 to 40 weeks
posttransplantation.BM cells were plated in methylcelluloseculture
as described in Materials and Methods. Eight to 10 colonies, containing greater than 5 x lo1 cells (which were mainly granulocyte/
macrophage and some multilineage),from each animal were individually analyzed by PCR indicated as an open or solid circle. Groups1
and 2 (GP 1 and Gp 2) were transplanted with 1 x l @and 8 x lo6
BM cells,respectively, from primary recipients transplanted with
rhG-CSF mobilized PBPC. Groups 3 and 4 IGp 3 and GP 4) were
transplanted with 1 x lp and 8 x l@ BM cells, respectively, from
primary recipients transplanted with rhG-CSF plus rrSCF mobilized
PBPC. Dashed line indicatesthe detection limit of 10% in the assay.
whole-blood PCR analysis, such as DNA template copy
number, amplification conditions, and the presence of cotransplanted male lymphocytes in circulation, which may be
long-lived invivo.",'6 Large colonies containing approximately greater than IO4 cells were individually picked from
cultures under an inverted microscope. Morphologic staining
of these colonies showed them to be mainly granulocytes
and macrophages, with about 10% of colonies being multilineage. DNA was prepared from each colony individually
and amplified with both Y-specific primers and PDGF B
receptor primers. Two hundred fifty colonies were analyzed
from these secondary recipients (Fig 2). Mice in groups 1,
2, and 3 had some animals that had Y-chromosome-positive
colonies. All mice (10 of 10)in group 4 had Y-chromosomepositive colonies, with 4 mice having 100% of colonies of
donor origin (Fig 2). These data showed a higher frequency
of long-term reconstituting cells from mice originally mobilized with rhG-CSF plus rrSCF compared with rhG-CSF
alone.
Cells in the lymphoid organs of the secondary recipient
mice were also analyzed for Y-chromosome-positive cells.
Ten mice from group 4 were killed at 6 months posttransplantation. DNA was extracted from spleen, thymus, and
lymph node and analyzed by semi-quantitative PCR (Fig 3).
All the lymph nodes, spleens, and thymuses had greater than
90%male-derived DNA. This showed that both myeloid and
lymphoid long-term repopulation originated from the rhGCSF plus rrSCF mobilized PBPC.
The BM cells from secondary recipients were transplanted
into irradiated tertiary female recipients. Ten secondary mice
from group 4 (Table 2) were killed at 26 weeks posttransplantation and the BM cells from the femurs and tibias of
each mouse were pooled and transplanted into 6 recipient
mice. Three mice received 1 X IO6 cells and 3 mice received
the remainder of the cells (range 3 to 10 X lo6, mean 6.9
t 1.7 X lo6 cells). At 24 weeks posttransplantation, the
tertiary recipients were analyzed for male cells in the peripheral blood. Figure 4 shows the analysis of tertiary recipient
mice that were transplanted with 1 X lo6BM cells. Nineteen
of 21 mice were male positive and 15 of 21 mice had greater
than 90% male-derived cells in the peripheral blood. All
animals transplanted with higher cell numbers similarly had
greater than 90% male-derived cells (data not shown).
DISCUSSION
In a previous report we showed that PBPC mobilized
by the combination of rhG-CSF plus rrSCF contained cells
capable of long-term reconstitution (>12 months) when
transplanted into irradiated recipients.* The data presented
in this study analyzed the potential of these mobilized PBPC
for reconstitution of secondary and tertiary recipients. A
number of studies have demonstrated that G-CSF or SCF
alone are able to mobilize cells that provide hematopoiesis
for 6 to 14 month^,'"'^ although higher doses of SCF were
needed to achieve the same mobilization as G-CSF. The
combination of rhG-CSF with a low dose of rrSCF clearly
showed synergy in PBPC mobilization?*20PBPC mobilized
by the combination of rhG-CSF plus rrSCF provided more
sustained engraftment than PBPC mobilized by rhG-CSF
alone. Previous studies by Harrison et a1 using BM cells
have shown that durability of LTF2A was related to the dose
ofBM
cells transplanted.8 Sustained donor engraftment
through serial transplantation is directly related to the number of stem cells present in the primary transplantation.
Therefore, the number of LTRC mobilized by rhG-CSF plus
rrSCF must be greater than the number of LTRC mobilized
by rhG-CSF alone.
We initially transplanted 2.5 X lo5 low-density cells
(equivalent to 20 pL whole blood) into primary recipients.
In the secondary transplantation, 1 X lo7 BM cells from
primary recipient were transplanted and demonstrated sustained engraftment in both lymphoid andmyeloid cells.
Therefore, one primary recipient mouse would be able to
reconstitute 20 secondary mice, assuming that there are 2.0
X 1
0' BM cells in a mouse. In the tertiary transplantation,
1 X lo6 BM cells from the secondary recipients were sufficient to give 90%male-derived blood nucleated cells. Thus,
one secondary recipient would be able to reconstitute 200
tertiary recipients. That is, 2.5 X 10' low-density cells from
the peripheral blood of rhG-CSF plus rrSCF treated mice
would be able to generate 4,000 tertiary recipients with 90%
of them having male-derived hematopoiesis. These results
suggest that rhG-CSF plus rrSCF mobilized PBPC contain
high numbers of LTRC.In addition, 2.5 X 10' peripheral
blood low-density cells have provided mature blood cells
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YAN ET AL
2306
% Male DNA loo 95 90 75 50 25 10 5 0
Standard
I
Mouse
I
1 2 3 4 5 6 7 8 9
1-1
Thymus
Lymph node
I
I-
PDGFBR
’
PDGFBR
Fig 3. Semi-quantitative PCR analysis of DNA
samples from spleen, thymus, and lymph node of
secondary recipient mice. Secondary recipients
transplanted with donor B cells were killed at 26
weeks posttransplantation. DNA was extracted from
spleens, thymuses, and lymph nodes. One hundred
nanograms of DNA was amplified with Y-specific
primer and PDGF B receptor primer. PCR-amplified
product was run on an agarose gel and Southern
blotted on t o a Nylon membrane. Membranes were
hybridized t o 32P-end-labeledSry or PDGF B receptor
internal oligos. The standard wasPCR products amplified from DNA extracted from spleens of a male
and a female mouse, respectively, and mixedas indicated. The figure shows the result
of nine mice (from
group 4 in Fig 2).
lm
Fig 4. PCR analysis of peripheral blood of21 tertiary recipient mice at 24 weeks posttransplantation. The tertiary recipient mice received
1.0 x 10’ BM cells from secondary recipients. PCR products were Southern blotted and hybridized with32P-labeledSry or PDGF B receptor
internal oligos.
for over 26 months, demonstrating extensive proliferative
potential throughthe serial transplantation. Whether the
LTRC undergo self-renewal is an interesting question and
would be consistent with the long-term engraftment generated through tertiary transplantation and potential engraftment of several thousand tertiary recipients.
The potential use of SCF in combination with G-CSF for
mobilization of PBPC has beenshown in several animal
models, including mouse, canine, and
These
studies demonstrated enhanced short-term engraftment of
irradiated animals transplanted with SCF plus G-CSF mobilized PBPC compared with G-CSF alone mobilized PBPC.
Based on these studies, clinical trials have been conducted
to compare mobilization of PBPC by rhG-CSF alone to mobilization with rhG-CSF plus rhSCF.’3”‘1In breast cancer
patients. the combination of rhG-CSF plus rhSCF at doses
of IO pg/kg/d or higher resulted in a twofold to threefold
increase in PBPC compared with rhG-CSF alone.” Transplantation of the PBPC after high-dose chemotherapy resulted in equivalent engraftment in bothpatient group~.’~
Additional clinical studies are required to determine if the
increase in PBPC leads to clinical benefits. The potential use
of mobilized PBPC for ex vivo expansion and gene therapy
will require optimal CD34 levels in PBPC samples and maximum numbers of cells with long-term engrafting potential.
The data presented in this study support the use of rhG-CSF
plus rhSCF mobilizing PBPC in these settings because they
would provide increased numbers of mobilized progenitors
and long-term engrafting cells.
ACKNOWLEDGMENT
The authors thank Dr G. Morstyn for continued support through
thesestudies, and J. Keysor forassistance inpreparation of the
figures.
REFERENCES
I . Molineux G, Pojda Z, Hampson IN, Dexter TM: Transplantation potential of peripheral blood stem cells induced by granulocyte
colony-stimulating factor. Blood 76:2153, 1990
2. Yan XQ, Briddell R, Hartley C, Stoney G , Samal B, McNiece
I: Mobilization of long-term hematopoietic reconstituting cells in
mice by the combination of stem cell factor plus granulocyte colonystimulating factor. Blood 84:795, 1994
3. Fibbe WE, Hamilton MS, Laterveer LL. Kibbelaar RE. Falkenburg JHF, Visser JWM. Willemze R: Sustained engraftment of mice
transplanted with IL-l-primed blood-derived stem cells. J lmmunol
148:417,1992
4. Briddell RA, Hartley CA, Smith KA, McNiece IK: Recombinant rat stem cell factor synergizes with recombinant human granulocyte colony-stimulating factor in vivo in mice to mobilize peripheral
blood progenitor cells that have enhanced repopulating potential.
Blood 82: 1720. 1993
From www.bloodjournal.org by guest on February 6, 2015. For personal use only.
MOBILIZATION OF LONG-TERM ENGRAFTINGCELLS
5. Ogden DA, Micklem HS: The fate of serially transplanted bone
marrow cell populations from young and olddonors. Transplantation
22:287, 1976
6. Brecher G, Neben S, Yee M, Bullis J, Cronkite EP: Pluripotential stem cells with normal or reduced self renewal survive lethal
irradiation. Exp Hematol 16:627, 1988
7. Harrison DE, Astle CM, Delaittre JA: Loss of proliferative
capacity in immunohematopoietic stem cells caused by serial transplantation rather than aging. J Exp Med 147:1526, 1978
8. Harrison DE, Stone M, Astle CM: Effects of transplantation
on the primitive immunohematopoietic stem cell. J Exp Med
172:431, 1990
9. Souza L, Boone T, Gabrilove J, Lai P, Zsebo K, Murdock D,
Cahzin V, Bruszewski J, Lu H, Chen K, Barendt J, Platzer E, Moore
M, Mertelsmann R, Welte K: Recombinant granulocyte colony-stimulating factor effects on normal and leukemic myeloid cells. Science
232:6 I , 1986
10. Martin FH, Suggs SV, Langley KE,Lu HS, Ting J, Okino
KH, Morris CF, McNiece IK, Jacobsen F W , Mendiaz EA, Birkett
NC, Smith KA, Johnson MJ, Parker VP, Flores JC, Patel AC, Fisher
EF, Erjavec HO, Herrera CJ, Wypych J, Sachdev RK, Pope JA,
Leslie I, Wen D, Lin CH, Cupples RL, Zsebo KM: Primary structure
and functional expression of rat and human stem cell factor DNAs.
Cell 63:203, 1990
1 I . Iscove NN: Culture of lymphocytes and hemopoietic cells in
serum-free medium, in Barnes D, Sirbascu D, Sat0 G (eds): Methods
for Serum-Free Culture of Neuronal and Lymphoid Cells. New York,
NY, Liss, 1984, p 169
12. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning. Cold
Spring Harbor, NY, Cold Spring Harbor Laboratory, 1989
13. Gubbay J, Vivian N, Economou A, Jackson D, Goodfellow
P, Lovell-Badge R: Inverted repeat structure of the Sry locus in
mice. Roc Natl Acad Sci USA 89:7953, 1992
14. Yarden Y, Escobedo JA, Kuang WJ, Yang-Feng TL, Daniel
TO, Tremble PM, Chen EY, Ando ME: Structure of the receptor
for platelet-derived growth factor helps define a family of closely
related growth factor receptors. Nature 323:226, 1986
15. Robinson SH, Brecher G, Lourie IS, HaleyJE: Leukocyte
labelling in rats during and after continuous infusion with tritidated
thymidine. Blood 26:28 I , 1965
16. Little JR, Brecher G, Bradley TF, Rose S: Determination of
lymphocyte turnover by continuous infusion of tritiated thymidine.
Blood 19:281, 1962
17. Neben S, Marcus K, Mauch P: Mobilization of hematopoietic
stem and progenitor cell subpopulations from the marrow to the
blood of mice following cyclophosphamide andor granulocyte colony-stimulating factor. Blood 8 1: 1960, 1993
18. Bodine DM, Seidel NE, Zsebo K M , Orlic D: In vivo adminis-
2307
tration of stem cell factor to mice increase the absolute number of
pluripotent hematopoietic stem cells. Blood 82:445, 1993
19. Bodine DM, Seidel NE, Orlic D, Nienhuis AW: Administration ofG-CSF and SCF to splenectomized mice increases the number
of peripheral blood stem cells which are efficiently transduced by
retroviruses. Blood 82:314a, 1993 (abstr, suppl I )
20. Andrews RG, Briddell R A , Knitter GH, Opie T, Bronsdon
M, Fausset P, Myerson D, Appelbaum FR. McNiece IK: Low dose
recombinant human stem cell factor (SCF) has a synergistic interaction with recombinant human granulocyte colony-stimulating factor
(G-CSF) in vivo for stimulating the circulation of progenitor cells
of multiple types in the peripheral blood of baboons. Blood 82:232a,
1993 (abstr, suppl 1)
21. De Revel T, Appelbaum F, Storb R, Schuening F, Nash R,
Deeg J, McNiece I, Andrews R, Graham T: Effects of granulocyte
colony-stimulating factor and stem cell factor, alone and incombination on the mobilization of peripheral blood cells that engraft lethally
irradiated dogs. Blood 83:3795, 1994
22. Andrews RG, Briddell RA, Knitter GH, Rowley SD, Appelbaum FR, McNiece IK: Rapid engraftment by peripheral blood progenitor cells mobilized by recombinant human stem cell factor and
recombinant human granulocyte colony-stimulating factor in nonhuman primates. Blood 85: 15, 1995
23. Briddell R, Glaspy J, Shpall EJ, LeMaistre F, Menchaca D,
McNiece I: Recombinant human stem cell factor (rhSCF) and filgrastim (rhG-CSF) synergize to mobilize myeloid, erythroid and
megakaryocyte progenitors in patients with breast cancer. Br J
Haematol 87:92, 1994 (suppl 1)
24. Glaspy J, McNiece I, LeMaistre F, Menchaca D, Briddell R,
Lill M, Jones R, Tami J, Morstyn G, Brown S, Shpall El: Effects
of stem cell factor (rhSCF) and filgrastim (rhG-CSF) on the mobilization of peripheral blood progenitor cells (PBPC) and hematological
recovery post transplant: prelimilary phase ID1 study results. Br J
Haematol 87:156, 1994 (suppl I )
25. Moskowitz C, Stiff P, Gordon M, Gabrilove J, Bayer R, Broun
R, Nichols C, Ho AD, Wyres M, Nimer S, McNiece I: The influence
of extensive prior chemotherapy on the mobilization of peripheral
blood progenitor cells (PBPC) using stem cell factor (rhSCF) and
filgrastim (rmetHuG-CSF) and on hematologic recovery post cyclophosphamide, BCNU, and VP-16 (CBV) in patients with relapsed
non-Hodgkin’s lymphoma (NHL): An interim analysis. Blood
84:107a, 1994 (abstr, suppl 1)
26. Begley CG, Basser R, Mansfield R, Maher D, To B, Juttner
c, Fox R, Cebon J, Grigg A, Szer J, McGrath K, Thomson B,
Sheridan W, Menchaca D, Collins J, Russell I, Green M: Randomized prospecrive study demonstrating a prolonged effect of SCF with
G-CSF (filgrastim) on PBPC in untreated patients: Early results.
Blood 84:25a, 1994 (abstr, suppl I )
From www.bloodjournal.org by guest on February 6, 2015. For personal use only.
1995 85: 2303-2307
Peripheral blood progenitor cells mobilized by recombinant human
granulocyte colony-stimulating factor plus recombinant rat stem cell
factor contain long-term engrafting cells capable of cellular
proliferation for more than two years as shown by serial
transplantation in mice
XQ Yan, C Hartley, P McElroy, A Chang, C McCrea and I McNiece
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