| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
(Stroke. 2001;32:1005.)
© 2001 American Heart Association, Inc.
Original Contributions |
From Henry Ford Health Sciences Center, Department of Neurology (J.C., Y.L., L.W., Z.Z., M.C.), Neurosurgery (D.L.), and Biostatistics and Research Epidemiology (M.L.), Detroit, Mich; and Oakland University, Department of Physics (M.C.), Rochester, Mich.
Correspondence to Michael Chopp, PhD, Henry Ford Hospital, Neurology Department, 2799 W Grand Blvd, Detroit, MI 48202. E-mail chopp{at}neuro.hfh.edu
| Abstract |
|---|
|
|
|---|
MethodsRats (n=32) were subjected to 2 hours of middle cerebral artery occlusion (MCAO). Test groups consisted of MCAO alone (group 1, n=6); intravenous infusion of 1x106 MSCs at 24 hours after MCAO (group 2, n=6); or infusion of 3x106 MSCs (group 3, n=7). Rats in groups 1 to 3 were euthanized at 14 days after MCAO. Group 4 consisted of MCAO alone (n=6) and group 5, intravenous infusion of 3x106 MSCs at 7 days after MCAO (n=7). Rats in groups 4 and 5 were euthanized at 35 days after MCAO. For cellular identification, MSCs were prelabeled with bromodeoxyuridine. Behavioral tests (rotarod, adhesive-removal, and modified Neurological Severity Score [NSS]) were performed before and at 1, 7, 14, 21, 28, and 35 days after MCAO. Immunohistochemistry was used to identify MSCs or cells derived from MSCs in brain and other organs.
ResultsSignificant recovery of somatosensory behavior and Neurological Severity Score (P<0.05) were found in animals infused with 3x106 MSCs at 1 day or 7 days compared with control animals. MSCs survive and are localized to the ipsilateral ischemic hemisphere, and a few cells express protein marker phenotypic neural cells.
ConclusionsMSCs delivered to ischemic brain tissue through an intravenous route provide therapeutic benefit after stroke. MSCs may provide a powerful autoplastic therapy for stroke.
Key Words: bone marrow transplantation middle cerebral artery occlusion neuronal plasticity stroke, experimental stromal cells rats
| Introduction |
|---|
|
|
|---|
Transplantation of adult MSCs directly into adult rat brain and spinal cord reduces functional deficits associated with stroke,10 11 traumatic brain injury, and spinal cord injury,12 respectively. MSCs express neural phenotype and migrate when placed in damaged brain10 11 and spinal cord.12 Systemic infusion of male BM cells into irradiated female mice results in an influx of Y chromosome cells into the brain over days to weeks and differentiation of these cells to microglia and astroglia.3 In addition, the male-derived BM cells systemically infused into female ischemic rats migrate preferentially to ischemic cortex.13 Thus, an alternative method to intracerebral MSC transplantation is to infuse these cells intravenously after in vitro expansion. Systemically infused MSCs can repopulate a number of nonhematopoietic tissues.6 Successful stromal chimerism has been achieved in murine systems with this approach.6 7 14 Koc et al15 have demonstrated the feasibility and safety of infusing culture-expanded autologous MSCs in patients with advanced breast cancer undergoing peripheral blood stem cell transplantation. In light of the utility of MSCs to treat neural injury and the potential vascular route of administration, in the present study, we test the hypothesis that intravenous infusion of MSCs from marrow reduces functional deficits after stroke in rats.
| Materials and Methods |
|---|
|
|
|---|
Experimental Groups
Experimental groups consist of group 1 (control):
rats given MCAO alone without donor cell administration (n=6); group 2:
rats given low-dose MSCs (1x106) injected
intravenously at 24 hours after MCAO (n=6); and group 3:
rats given high-dose MSCs (3x106) injected
intravenously at 24 hours after MCAO (n=7). The animals of
groups 1, 2, and 3 were killed at 14 days after MCAO. To test the
effects of delayed (7-day) treatment, we included 2 additional groups:
Group 4 rats (control) were given MCAO alone without donor cell
administration (n=6) and were killed at 35 days after MCAO; group 5
rats were given high-dose MSCs (3x106)
injected intravenously at 7 days after MCAO and were killed
at 35 days (n=7) after MCAO. The selection of an extended survival time
(35 days) was based on the supposition that late treatment, 7 days
after MCAO, provides a delayed functional
benefit.
Transplantation Procedures
Primary cultures of BM cells were obtained 48 hours
after treating donor rats with 5-fluorouracil (150 mg/kg); MSCs were
separated, as previously
described.3 11 All
transplantation procedures were performed under aseptic conditions. At
1 or 7 days after ischemia, randomly selected animals received
transplantation. Animals were anesthetized with 3.5% halothane
and then maintained with 1.0% to 2.0% halothane in 70%
N2O and 30% O2 by a face
mask mounted in a Kopf stereotaxic frame (model 51603,
Stoelting Co). Approximately 1x106 or
3x106 MSCs in 1 mL total fluid volume were
injected into a tail vein. Immunosuppressants were not used in any
animal.
Behavioral Testing
In all animals, a battery of behavioral tests was
performed before MCAO and at 1, 7, 14, 21, 28, and 35 days after MCAO
by an investigator who was blinded to the experimental groups. For the
rotarod test,18 19 rats were placed on an accelerating rotarod cylinder, and
the time the animals remained on the rotarod was measured. The speed
was slowly increased from 4 to 40 rpm within 5 minutes. A trial ended
if the animal fell off the rungs or gripped the device and spun around
for 2 consecutive revolutions without attempting to walk on the rungs.
The animals were trained 3 days before MCAO. The mean duration (in
seconds) on the device was recorded with 3 rotarod measurements 1
day before surgery. Motor test data are presented as percentage
of mean duration (3 trials) on the rotarod compared with the internal
baseline control (before surgery).
For the adhesive-removal somatosensory test,18 20 21 somatosensory deficit was measured both before and after surgery. All rats were familiarized with the testing environment. In the initial test, 2 small pieces of adhesive-backed paper dots (of equal size, 113.1 mm2) were used as bilateral tactile stimuli occupying the distal-radial region on the wrist of each forelimb. The rat was then returned to its cage. The time to remove each stimulus from forelimbs was recorded on 5 trials per day. Individual trials were separated by at least 5 minutes. Before surgery, the animals were trained for 3 days. Once the rats were able to remove the dots within 10 seconds, they were subjected to MCAO.
Table 1
shows a set of modified Neurological
Severity Scores
(NSS).22 23 24 25
Neurological function was graded on a scale of 0 to 18 (normal score,
0; maximal deficit score, 18). NSS is a composite of motor, sensory,
reflex, and balance tests.26
In the severity scores of injury, 1 score point is awarded for the
inability to perform the test or for the lack of a tested reflex; thus,
the higher score, the more severe is the injury.
|
Histological and
Immunohistochemical Assessment
Animals were allowed to survive for 14 or 35 days
after MCAO, and at that time animals were reanaesthetized with
ketamine (44 mg/kg) and xylazine (13 mg/kg). Rat brains were
fixed by transcardial perfusion with saline, followed by perfusion and
immersion in 4% paraformaldehyde, and the brain,
heart, liver, spleen, lung, kidney, and muscle were embedded in
paraffin. The cerebral tissues were cut into 7 equally spaced (2
mm) coronal blocks. A series of adjacent 6-µm-thick sections were cut
from each block in the coronal plane and were stained with hematoxylin
and eosin. The 7 brain sections were traced by the Global Laboratory
Image analysis system (Data Translation). The indirect lesion
area, in which the intact area of the ipsilateral hemisphere was
subtracted from the area of the contralateral hemisphere, was
calculated.27 The lesion
volume is presented as a volume percentage of the lesion
compared with the contralateral hemisphere.
Single and double immunohistochemical staining28 was used to identify cells derived from MSCs. Briefly, a standard paraffin block was obtained from the center of the lesion, corresponding to coronal coordinates for bregma -1~1 mm. A series of 6-µm-thick sections at various levels (100-µm interval) were cut from this block and were analyzed by light and fluorescent microscopy (Olympus, BH-2). To detect the distribution of transplanted MSCs in other organs (ie, heart, liver, lung, spleen, kidney, muscle, and bone marrow), 3 sections (6 µm thick, 100-µm interval) from each organ were obtained and numbers of bromodeoxyuridine (BrdU)-reactive cells measured. Measurements of BrdU-reactive cells in organs other than brain were performed in all rats subjected to 1-day or 7-day treatment with 3x106 MSCs. After deparaffinization, sections were placed in boiled citrate buffer (pH 6.0) within a microwave oven (650 to 720 W). After blocking in normal serum, sections were treated with the monoclonal antibody against BrdU (Calbiochem) diluted at 1:100 in PBS. After sequential incubation with peroxidase-conjugated rabbit anti-mouse IgG (dilution 1:100; Dakopatts), the secondary antibody was bound to the first antibody against BrdU. Diaminobenzidine (DAB) was then used as a chromogen for light microscopy. Counterstaining of sections by hematoxylin was also performed. Cells derived from MSCs were identified by morphological criteria and by immunohistochemical staining with BrdU (the tracer) present in the nuclei of donor cells. BrdU found in the parenchymal cells or in the cytoplasm of macrophage-like cells was not counted. Analysis of BrdU-positive cells is based on the evaluation of an average of 10 histology slides of brain. All BrdU-reactive cells, with BrdU clearly localized to the nucleus, were counted throughout all 10 coronal sections. For information on the relative presence of MSCs within other organs, 3 slides from each organ were obtained from each experimental animal, and an estimate was made of the percentage of BrdU-positive cells to endogenous organ-specific cells.
To visualize the cellular colocalization of BrdU-specific and cell-typespecific markers in the same cells, double staining was used. Brain sections were treated with cell-typespecific antibodies, a neuronal nuclear antigen (NeuN for neurons, dilution 1:200; Chemicon), microtubule-associated protein 2 (MAP-2 for neurons, dilution 1:200; Boehringer Mannheim), and glial fibrillary acidic protein (GFAP for astrocytes, dilution 1:1000; Dako). Each coronal section was first treated with the primary BrdU monoclonal antibody, as described above. FITC-conjugated antibody (Calbiochem) was used for double-label immunoreactivity identification. Negative control sections from each animal received identical preparations for immunohistochemical staining, except that primary antibodies were omitted. A total of 500 BrdU-positive cells per animal from multiple adjacent (100-µm interval) sections were counted to obtain the percentage of BrdU cells colocalized with cell-typespecific markers (NeuN, MAP2, and GFAP) by double staining.
Laser Scanning Confocal Microscopy
Colocalization of BrdU with neuronal marker was
conducted by laser scanning confocal microscopy (LSCM) with the use of
a Bio-Rad MRC 1024 (argon and krypton) laser-scanning confocal imaging
system mounted onto a Zeiss microscope
(Bio-Rad).29 For
immunofluorescence double-labeled coronal sections,
green (FITC for BrdU) and red cyanine-5.18 (Cy5 for MAP-2 or NeuN)
fluorochromes on the sections were excited by a laser beam at 488 nm
and 647 nm; emissions were sequentially acquired with 2 separate
photomultiplier tubes through 522 nm and 680 nm emission filters,
respectively. Areas of interest were scanned with a x40 oil immersion
objective lens in 260.6x260.6-µm format in the
x-y direction and 0.5 µm in
the z
direction.
Statistical Analysis
The behavior scores (rotarod test, adhesive-removal
test, and NSS), were evaluated for normality. Repeated-measures
analysis was conducted to test the treatment effect on the
behavior score. The analysis began with testing for the
treatment-time interaction at the significance level 0.1, then testing
for the overall treatment effect if there was no interaction detected
at the 0.05 level. A subgroup analysis of the treatment effect
on each behavior score at each time was conducted at the 0.05
significance level if a treatment-time interaction at the 0.1 level or
an overall treatment effect at the 0.05 level was found. Otherwise,
subgroup analyses would be considered as exploratory. The means
(SD) and probability value for testing the difference between treated
and control groups are presented.
For the 2 control groups, 1 control group had complete behavioral scoring up to 14 days after ischemia before they were euthanized for histological analysis; the other control group had complete behavioral scores up to 35 days after ischemia. The control animals were shared for testing the treatment effect with different doses and treatment given at different times after ischemia.
| Results |
|---|
|
|
|---|
|
|
Histology
The blood gasses were within normal ranges for all
animals and did not differ among groups (data not shown). Within the
6-µm-thick coronal sections stained with hematoxylin and eosin, dark
and red neurons were observed in the ischemic core of all rats
subjected to MCAO with and without donor transplantation at 14 and 35
days after MCAO. No significant reduction of volume of ischemic
damage was detected in rats with donor treatment compared with control
rats subjected to MCAO alone
(Table 2
).
|
Within the brain tissue, cells derived from MSCs were
characterized by round-to-oval dark brown nuclei with irregularly
shaped and thin cytoplasm by BrdU staining. Only cells with this
morphology and with BrdU localized solely to nucleus were counted as
MSCs
(Figure 3a
). MSCs identified by BrdU immunoreactivity
survived and were distributed throughout the damaged brain of recipient
rats. Some cells, the vast majority within the lesion, contained BrdU
within the cytoplasm
(Figure 3b
). These cells are considered as
macrophages and are not counted as MSCs. The number of
BrdU-reactive cells detected from an average of 10 histology slides per
MSC transplantation animal is given in
Table 2
. Higher levels BrdU-reactive cells were seen in the
brain at high-dose (3x106) MSC
transplantation group than in the low-dose
(1x106) MSC group at 24 hours after MCAO
(P<0.01)
(Table 2
). BrdU-reactive cells were observed in multiple
areas of the ipsilateral hemisphere, including cortexes, striatum of
the ipsilateral hemisphere. The vast majority of BrdU-labeled MSCs were
located in the ischemic core and its boundary zone. Few cells
were observed in the contralateral hemisphere.
|
In organs other than brain, as an approximate percentage of
endogenous organ-specific cells, scattered BrdU-positive
cells were detected in bone marrow (
2% to 4%) and in muscle,
spleen, kidney, lung, and liver (
0.01% to 0.5%)
(Figure 3, c and d
). Most BrdU-positive cells encircle vessels
in these organs, with few cells located in parenchyma.
Double-staining immunohistochemistry of brain sections
revealed that some BrdU-positive cells were reactive for the neuronal
markers NeuN
(Figure 3, e and f
) and MAP-2 and for the astrocyte marker
GFAP
(Figure 3, g and h
). The percentage of BrdU that labeled
expressed NeuN, MAP-2, and GFAP proteins was
1%,
2%, and
5%, respectively.
Figure 4
shows LSCM images from the coronal sections
immunofluorescently stained with antibodies against MAP2, BrdU,
and NeuN. Colocalization of immunofluorescent labels for MAP2
and BrdU
(Figure 4, A to G
) and for NeuN and BrdU
(Figure 4, H to J
) were
present.
|
| Discussion |
|---|
|
|
|---|
What are the mechanisms or factors that promote reduced deficits with MSC transplantation after stroke? One possibility is that the MSCs integrate into the tissue, replace damaged cells, and reconstruct neural circuitry. However, we have no clear evidence that the MSCs function in this way, and although some cells express neural cell phenotype, we have no evidence that these cells develop contacts with other neurons. Reconstruction of neural circuitry is not always a prerequisite for functional recovery.30 A more reasonable hypothesis is that interaction of MSCs with the host brain may lead to production of trophic factors,31 which may contribute to recovery of function lost as a result of lesions, the mechanisms of which are unidentified.32 MSCs constitutively secrete interleukins (IL)-6, IL-7, IL-8, IL-11, IL-12, IL-14, IL-15, macrophage colony-stimulating factor, Flt-3 ligand, and stem-cell factor.33 34 These cytokines are survival, growth, and/or differentiation factors for murine hippocampal neuronal progenitor cells.35 36 37 38 39 MSCs also contain catecholamines and may release specific neurotransmitters.40 Thus, reduction of ischemic-induced deficits by MSC transplantation may be due to the production of trophic factors by MSCs.
Nonhematopoietic BM stroma is composed of mesenchymal cells, including fibroblasts, osteoblasts, and adipocytes, in addition to endothelial cells.1 Most nonhematopoietic stromal progenitor cells appear to be more consistent with an endothelial rather than a fibroblast cell origin.41 42 In response to severe ischemia or cytokine stimuli, stromal progenitors may expand and be recruited along with endothelia progenitor cells (EPCs) and consequently contribute to neovascularization and/or wound-healing processes.43 In the present study, BrdU-labeled cells encircled vessels of organs at 14 days after injection. We speculate that transplanted MSCs may function as EPC. EPC mobilization may ultimately represent a potential strategy for clinical therapy of ischemic vascular disease.
More MSCs were found in the lesioned hemisphere than in the intact hemisphere. These data are consistent with reports that male BM cells systemically infused into female ischemic rats migrate preferentially to ischemic cortex.13 The mechanisms responsible for intravenously infused BM migration into brain and its intraparenchymal distribution are not clear. Disruption of the blood-brain barrier may facilitate selective entry of MSCs into ischemic brain compared with nonischemic contralateral cerebral tissue. In addition, there are other mechanisms that may promote migration of MSCs into brain. Approximately 20% of microglia are thought to originate from the marrow.44 45 MSCs intravenously injected into irradiated mice continue to replicate in vivo, and over a period of weeks they populate several connective tissues including bone, cartilage, lung, and brain.6 By systemic administration, BM-derived myogenic progenitors migrate into a degenerating muscle and participate in the regeneration process.46 These cells appear to be recruited by long-range, possibly inflammatory, signals originating from the degenerating tissue, and they probably access the damaged muscle from the circulation, together with granulocytes and macrophages.46 Natural migration of BM cells from one hematopoietic microenvironment to the other may occur.47 This movement is genetically controlled in part through changes in expression of cell surface adhesion molecules, such as intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and neural cell adhesion molecule.48 49
In our previous study, we transplanted BM or MSCs cells into ischemic brain,10 11 18 and these cells migrate, differentiate, and reduce functional deficits after stroke.11 18 Injection of devitalized BrdU-labeled MSCs directly into brain resulted in no improvement in functional outcome, and outcome was no different from that detected in rats with intracerebral PBS injection. Local intracerebral injection induces local brain damage,50 and particularly, multiple injections may not be clinically acceptable. The most important finding of this study is that BM-derived MSCs delivered to ischemic tissue through an intravenous route provide therapeutic benefit. This simple approach for cell therapy, which does not necessitate invasive stereotaxic operations, could potentially target pathological sites in a number of brain disorders. Logistical and ethical concerns about the use of fetal cells for transplantation therapy can be eliminated by exploiting MSCs as an alternative autologous graft source.
| Acknowledgments |
|---|
Received August 21, 2000; revision received November 14, 2000; accepted December 19, 2000.
| References |
|---|
|
|
|---|
2.
Pittenger MF,
Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA,
Simonetti DW, Craig S, Marshak DR. Multilineage potential of adult
human mesenchymal stem cells.
Science. 1999;284:143147.
3.
Eglitis MA, Mezey
E. Hematopoietic cells differentiate into both microglia and macroglia
in the brains of adult mice. Proc Natl
Acad Sci
U S A. 1997;94:40804085.
4. Ashton BA, Allen TD, Howlett CR, Eaglesom CC, Hattori A, Owen M. Formation of bone and cartilage by marrow stromal cells in diffusion chambers in vivo. Clin Orthop. 1980;151:294307.
5.
Bennett JH, Joyner
CJ, Triffitt JT, Owen ME. Adipocytic cells cultured from marrow have
osteogenic potential. J Cell
Sci. 1991;99:131139.
6.
Pereira RF, Halford
KW, OHara MD, Leeper DB, Sokolov BP, Pollard MD, Bagasra O, Prockop
DJ. Cultured adherent cells from marrow can serve as long-lasting
precursor cells for bone, cartilage, and lung in irradiated mice.
Proc Natl Acad Sci
U S A. 1995;92:48574861.
7.
Pereira RF, OHara
MD, Laptev AV, Halford KW, Pollard MD, Class R, Simon D, Livezey K,
Prockop DJ. Marrow stromal cells as a source of progenitor cells for
nonhematopoietic tissues in transgenic mice with a phenotype of
osteogenesis imperfecta. Proc Natl Acad
Sci
U S A. 1998;95:11421147.
8. Prockop DJ. Heritable osteoarthritis: diagnosis and possible modes of cell and gene therapy. Osteoarthritis Cartilage. 1999;7:364366.[Medline] [Order article via Infotrieve]
9.
Kopen GC, Prockop
DJ, Phinney DG. Marrow stromal cells migrate throughout forebrain and
cerebellum, and they differentiate into astrocytes after injection into
neonatal mouse brains. Proc Natl Acad Sci
U S A. 1999;96:1071110716.
10. Li Y, Chen J, Chopp M. Adult bone marrow transplantation after stroke in adult rats. Cell Transplant. In press.
11. Li Y, Chopp M, Chen J, Wang L, Gautam SC, Xu Y, Zhang ZG. Interastriatal transplantation of bone marrow stromal cells (MSCs) improves functional recovery after stroke in adult mice. J Cereb Blood Flow Metab.. 2000;20:13111319.[Medline] [Order article via Infotrieve]
12. Chopp M, Zhang XH, Li Y, Wang L, Chen J, Lu D, LU M, Rosenblum M. Spinal cord injury in rat: treatment with bone marrow stromal cell transplantation. Neuroreport.. 2000;11:30013005.[Medline] [Order article via Infotrieve]
13. Eglitis MA, Dawson D, Park KW, Mouradian MM. Targeting of marrow-derived astrocytes to the ischemic brain. Neuroreport. 1999;10:12891292.[Medline] [Order article via Infotrieve]
14. Huss R, Smith FO, Myerson DH, Deeg HJ. Homing and immunogenicity of murine stromal cells transfected with xenogeneic MHC class II genes. Cell Transplant. 1995;4:483491.[Medline] [Order article via Infotrieve]
15. Koc ON, Peters C, Aubourg P, Raghavan S, Dyhouse S, DeGasperi R, Kolodny EH, Yoseph YB, Gerson SL, Lazarus HM, Caplan AI, Watkins PA, Krivit W. Bone marrow-derived mesenchymal stem cells remain host-derived despite successful hematopoietic engraftment after allogeneic transplantation in patients with lysosomal and peroxisomal storage diseases. Exp Hematol. 1999;27:16751681.[Medline] [Order article via Infotrieve]
16. Chen H, Chopp M, Zhang ZG, Garcia JH. The effect of hypothermia on transient middle cerebral artery occlusion in the rat. J Cereb Blood Flow Metab. 1992;12:621628.[Medline] [Order article via Infotrieve]
17.
Longa EZ,
Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery
occlusion without craniectomy in rats.
Stroke. 1989;20:8491.
18. Chen J, Li Y, Chopp M. Intracerebral transplantation of bone marrow with BDNF after MCAo in rat. Neuropharmacology. 2000;39:711716.[Medline] [Order article via Infotrieve]
19. Hamm RJ, Pike BR, ODell DM, Lyeth BG, Jenkins LW. The rotarod test: an evaluation of its effectiveness in assessing motor deficits following traumatic brain injury. J Neurotrauma. 1994;11:187196.[Medline] [Order article via Infotrieve]
20. Schallert T, Hernandez TD, Barth TM. Recovery of function after brain damage: severe and chronic disruption by diazepam. Brain Res. 1986;379:104111.[Medline] [Order article via Infotrieve]
21. Zhang L, Chen J, Li Y, Zhang ZG, Chopp M. Quantitative measurement of motor and somatosensory impairments mild (30 min) and severe (2 h) transient middle cerebral artery occlusion in rats. J Neurol Sci. 2000;174:141146.[Medline] [Order article via Infotrieve]
22. Borlongan CV, Randall TS, Cahill DW, Sanberg PR. Asymmetrical motor behavior in rats with unilateral striatal excitotoxic lesions as revealed by the elevated body swing test. Brain Res. 1995;676:231234.[Medline] [Order article via Infotrieve]
23. Shohami E, Novikov M, Bass R. Long-term effect of HU-211, a novel non-competitive NMDA antagonist, on motor and memory functions after closed head injury in the rat. Brain Res. 1995;674:5562.[Medline] [Order article via Infotrieve]
24. Schallert T, Kozlowski DA, Humm JL, Cocke RR. Use-dependent structural events in recovery of function. Adv Neurol. 1997;73:229238.[Medline] [Order article via Infotrieve]
25. Chen Y, Constantini S, Trembovler V, Weinstock M, Shohami E. An experimental model of closed head injury in mice: pathophysiology, histopathology, and cognitive deficits. J Neurotrauma. 1996;13:557568.[Medline] [Order article via Infotrieve]
26. Germano AF, Dixon CE, dAvella D, Hayes RL, Tomasello F. Behavioral deficits following experimental subarachnoid hemorrhage in the rat. J Neurotrauma. 1994;11:345353.[Medline] [Order article via Infotrieve]
27. Swanson RA, Morton MT, Tsao-Wu G, Savalos RA, Davidson C, Sharp FR. A semiautomated method for measuring brain infarct volume. J Cereb Blood Flow Metab. 1990;10:290293.[Medline] [Order article via Infotrieve]
28. Li Y, Powers C, Jiang N, Chopp M. Intact, injured, necrotic and apoptotic cells after focal cerebral ischemia in the rat. J Neurol Sci. 1998;156:119132.[Medline] [Order article via Infotrieve]
29.
Zhang ZG, Chopp
M, Goussev A, Lu D, Morris D, Tsang W, Powers C, Ho KL. Cerebral
microvascular obstruction by fibrin is associated with upregulation of
PAI-1 acutely after onset of focal embolic ischemia in rats.
J Neurosci. 1999;19:1089810907.
30. Dunnett SB. Behavioural consequences of neural transplantation. J Neurol. 1994;242(suppl 1):S43S53.
31. Unsicker K. Growth factors in Parkinsons disease. Prog Growth Factor Res. 1994;5:7387.[Medline] [Order article via Infotrieve]
32. Hefti F. Nerve growth factor promotes survival of septal cholinergic neurons after fimbrial transections. J Neurosci. 1986;6:21552162.[Abstract]
33. Majumdar MK, Thiede MA, Mosca JD, Moorman M, Gerson SL. Phenotypic and functional comparison of cultures of marrow-derived mesenchymal stem cells (MSCs) and stromal cells. J Cell Physiol. 1998;176:5766.[Medline] [Order article via Infotrieve]
34.
Eaves CJ, Cashman
JD, Kay RJ, Dougherty GJ, Otsuka T, Gaboury LA, Hogge DE, Lansdorp PM,
Eaves AC, Humphries RK. Mechanisms that regulate the cell cycle status
of very primitive hematopoietic cells in long-term human marrow
cultures, II: analysis of positive and negative regulators
produced by stromal cells within the adherent layer.
Blood. 1991;78:110117.
35. Suzumura A. The cytokines and their functions on neural cells. Tanpakushitsu Kakusan Koso. 1995;40:691699.[Medline] [Order article via Infotrieve]
36. Mehler MF, Rozental R, Dougherty M, Spray DC, Kessler JA. Cytokine regulation of neuronal differentiation of hippocampal progenitor cells. Nature. 1993;362:6265.[Medline] [Order article via Infotrieve]
37. Maysinger D, Berezovskaya O, Fedoroff S. The hematopoietic cytokine colony stimulating factor 1 is also a growth factor in the CNS, II: microencapsulated CSF-1 and LM-10 cells as delivery systems. Exp Neurol. 1996;141:4756.[Medline] [Order article via Infotrieve]
38. Berezovskaya O, Maysinger D, Fedoroff S. The hematopoietic cytokine, colony-stimulating factor 1, is also a growth factor in the CNS: congenital absence of CSF-1 in mice results in abnormal microglial response and increased neuron vulnerability to injury. Int J Dev Neurosci. 1995;13:285299.[Medline] [Order article via Infotrieve]
39. Berezovskaya O, Maysinger D, Fedoroff S. Colony stimulating factor-1 potentiates neuronal survival in cerebral cortex ischemic lesion. Acta Neuropathol (Berl). 1996;92:479486.[Medline] [Order article via Infotrieve]
40. Maestroni GJ, Cosentino M, Marino F, Togni M, Conti A, Lecchini S, Frigo G. Neural and endogenous catecholamines in the bone marrow: circadian association of norepinephrine with hematopoiesis? Exp Hematol. 1998;26:11721177.[Medline] [Order article via Infotrieve]
41.
Perkins S,
Fleischman RA. Stromal cell progeny of murine bone marrow fibroblast
colony-forming units are clonal endothelial-like cells
that express collagen IV and laminin.
Blood. 1990;75:620625.
42. Fleischman RA, Simpson F, Gallardo T, Jin XL, Perkins S. Isolation of endothelial-like stromal cells that express Kit ligand and support in vitro hematopoiesis. Exp Hematol. 1995;23:14071416.[Medline] [Order article via Infotrieve]
43. Takahashi T, Kalka C, Masuda H, Chen D, Silver M, Kearney M, Magner M, Isner JM, Asahara T. Ischemia- and cytokine-induced mobilization of bone marrow-derived endothelial progenitor cells for neovascularization. Nat Med. 1999;5:434438.[Medline] [Order article via Infotrieve]
44.
Krall WJ,
Challita PM, Perlmutter LS, Skelton DC, Kohn DB. Cells expressing human
glucocerebrosidase from a retroviral vector repopulate
macrophages and central nervous system microglia after murine
bone marrow transplantation.
Blood. 1994;83:27372748.
45.
Kennedy DW,
Abkowitz JL. Kinetics of central nervous system microglial and
macrophage engraftment: analysis using a transgenic
bone marrow transplantation model.
Blood. 1997;90:986993.
46.
Ferrari G,
Cusella-De Angelis G, Coletta M, Paolucci E, Stornaiuolo A, Cossu G,
Mavilio F. Muscle regeneration by bone marrow-derived myogenic
progenitors. Science. 1998;279:15281530.
47.
Vermeulen M, Le
Pesteur F, Gagnerault MC, Mary JY, Sainteny F, Lepault F. Role of
adhesion molecules in the homing and mobilization of murine
hematopoietic stem and progenitor cells.
Blood. 1998;92:894900.
48. Haraldsen G, Kvale D, Lien B, Farstad IN, Brandtzaeg P. Cytokine-regulated expression of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) in human microvascular endothelial cells. J Immunol. 1996;156:25582565.[Abstract]
49.
Quesenberry PJ,
Becker PS. Stem cell homing: rolling, crawling, and nesting.
Proc Natl Acad Sci
U S A. 1998;95:1515515157.
50.
Gage FH, Coates
PW, Palmer TD, Kuhn HG, Fisher LJ, Suhonen JO, Peterson DA, Suhr ST,
Ray J. Survival and differentiation of adult neuronal progenitor
cells transplanted to the adult brain.
Proc Natl Acad Sci
U S A. 1995;92:1187911883.
This article has been cited by other articles:
![]() |
The STEPS Participants Stem Cell Therapies as an Emerging Paradigm in Stroke (STEPS): Bridging Basic and Clinical Science for Cellular and Neurogenic Factor Therapy in Treating Stroke Stroke, February 1, 2009; 40(2): 510 - 515. [Abstract] [Full Text] [PDF] |
||||
![]() |
Z. Liu, Y. Li, X. Zhang, S. Savant-Bhonsale, and M. Chopp Contralesional Axonal Remodeling of the Corticospinal System in Adult Rats After Stroke and Bone Marrow Stromal Cell Treatment Stroke, September 1, 2008; 39(9): 2571 - 2577. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. A. Knight, Y. Han, T. N. Nagaraja, P. Whitton, J. Ding, M. Chopp, and D. M. Seyfried Temporal MRI Assessment of Intracerebral Hemorrhage in Rats Stroke, September 1, 2008; 39(9): 2596 - 2602. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. D. Muller, K. M. Hanumanthiah, K. Diederich, S. Schwab, W.-R. Schabitz, and C. Sommer Brain-Derived Neurotrophic Factor But Not Forced Arm Use Improves Long-Term Outcome After Photothrombotic Stroke and Transiently Upregulates Binding Densities of Excitatory Glutamate Receptors in the Rat Brain Stroke, March 1, 2008; 39(3): 1012 - 1021. [Abstract] [Full Text] [PDF] |
||||
![]() |
X. Cui, J. Chen, A. Zacharek, Y. Li, C. Roberts, A. Kapke, S. Savant-Bhonsale, and M. Chopp Nitric Oxide Donor Upregulation of Stromal Cell-Derived Factor-1/Chemokine (CXC Motif) Receptor 4 Enhances Bone Marrow Stromal Cell Migration into Ischemic Brain After Stroke Stem Cells, November 1, 2007; 25(11): 2777 - 2785. [Abstract] [Full Text] [PDF] |
||||
![]() |
G. Chamberlain, J. Fox, B. Ashton, and J. Middleton Concise Review: Mesenchymal Stem Cells: Their Phenotype, Differentiation Capacity, Immunological Features, and Potential for Homing Stem Cells, November 1, 2007; 25(11): 2739 - 2749. [Abstract] [Full Text] [PDF] |
||||
![]() |
E. Shi, T. Kazui, X. Jiang, N. Washiyama, K. Yamashita, H. Terada, and A. H. M. Bashar Therapeutic Benefit of Intrathecal Injection of Marrow Stromal Cells on Ischemia-Injured Spinal Cord Ann. Thorac. Surg., April 1, 2007; 83(4): 1484 - 1490. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. De Becker, P. Van Hummelen, M. Bakkus, I. Vande Broek, J. De Wever, M. De Waele, and I. Van Riet Migration of culture-expanded human mesenchymal stem cells through bone marrow endothelium is regulated by matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-3 Haematologica, April 1, 2007; 92(4): 440 - 449. [Abstract] [Full Text] [PDF] |
||||
![]() |
S. Jomura, M. Uy, K. Mitchell, R. Dallasen, C. J. Bode, and Y. Xu Potential Treatment of Cerebral Global Ischemia with Oct-4+ Umbilical Cord Matrix Cells Stem Cells, January 1, 2007; 25(1): 98 - 106. [Abstract] [Full Text] [PDF] |
||||
![]() |
C. Buhnemann, A. Scholz, C. Bernreuther, C. Y. Malik, H. Braun, M. Schachner, K. G. Reymann, and M. Dihne Neuronal differentiation of transplanted embryonic stem cell-derived precursors in stroke lesions of adult rats Brain, December 1, 2006; 129(12): 3238 - 3248. [Abstract] [Full Text] [PDF] |
||||
![]() |
N. Sprigg, P. M. Bath, L. Zhao, M. R. Willmot, L. J. Gray, M. F. Walker, M. S. Dennis, and N. Russell Granulocyte-Colony-Stimulating Factor Mobilizes Bone Marrow Stem Cells in Patients With Subacute Ischemic Stroke: The Stem Cell Trial of Recovery EnhanceMent After Stroke (STEMS) Pilot Randomized, Controlled Trial (ISRCTN 16784092) Stroke, December 1, 2006; 37(12): 2979 - 2983. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. M. Coyne, A. J. Marcus, D. Woodbury, and I. B. Black Marrow Stromal Cells Transplanted to the Adult Brain Are Rejected by an Inflammatory Response and Transfer Donor Labels to Host Neurons and Glia Stem Cells, November 1, 2006; 24(11): 2483 - 2492. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. Liu, O. Honmou, K. Harada, K. Nakamura, K. Houkin, H. Hamada, and J. D. Kocsis Neuroprotection by PlGF gene-modified human mesenchymal stem cells after cerebral ischaemia Brain, October 1, 2006; 129(10): 2734 - 2745. [Abstract] [Full Text] [PDF] |
||||
![]() |
E. Pacary, H. Legros, S. Valable, P. Duchatelle, M. Lecocq, E. Petit, O. Nicole, and M. Bernaudin Synergistic effects of CoCl2 and ROCK inhibition on mesenchymal stem cell differentiation into neuron-like cells J. Cell Sci., July 1, 2006; 119(13): 2667 - 2678. [Abstract] [Full Text] [PDF] |
||||
![]() |
E. Shi, T. Kazui, X. Jiang, N. Washiyama, K. Yamashita, H. Terada, and A. H. M. Bashar Intrathecal Injection of Bone Marrow Stromal Cells Attenuates Neurologic Injury After Spinal Cord Ischemia Ann. Thorac. Surg., June 1, 2006; 81(6): 2227 - 2234. [Abstract] [Full Text] [PDF] |
||||
![]() |
A. Y. Khakoo, S. Pati, S. A. Anderson, W. Reid, M. F. Elshal, I. I. Rovira, A. T. Nguyen, D. Malide, C. A. Combs, G. Hall, et al. Human mesenchymal stem cells exert potent antitumorigenic effects in a model of Kaposi's sarcoma J. Exp. Med., May 15, 2006; 203(5): 1235 - 1247. [Abstract] [Full Text] [PDF] |
||||
![]() |
C. M. Sheridan, D. Rice, P. S. Hiscott, D. Wong, and D. L. Kent The Presence of AC133-Positive Cells Suggests a Possible Role of Endothelial Progenitor Cells in the Formation of Choroidal Neovascularization. Invest. Ophthalmol. Vis. Sci., April 1, 2006; 47(4): 1642 - 1645. [Abstract] [Full Text] [PDF] |
||||
![]() |
Y. Ma, Y. Xu, Z. Xiao, W. Yang, C. Zhang, E. Song, Y. Du, and L. Li Reconstruction of Chemically Burned Rat Corneal Surface by Bone Marrow-Derived Human Mesenchymal Stem Cells Stem Cells, February 1, 2006; 24(2): 315 - 321. [Abstract] [Full Text] [PDF] |
||||
![]() |
L. Zhang, R. L. Zhang, Y. Wang, C. Zhang, Z. G. Zhang, H. Meng, and M. Chopp Functional Recovery in Aged and Young Rats After Embolic Stroke: Treatment With a Phosphodiesterase Type 5 Inhibitor Stroke, April 1, 2005; 36(4): 847 - 852. [Abstract] [Full Text] [PDF] |
||||
![]() |
K. Hanabusa, N. Nagaya, T. Iwase, T. Itoh, S. Murakami, Y. Shimizu, W. Taki, K. Miyatake, and K. Kangawa Adrenomedullin Enhances Therapeutic Potency of Mesenchymal Stem Cells After Experimental Stroke in Rats Stroke, April 1, 2005; 36(4): 853 - 858. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. Chen, A. Zacharek, C. Zhang, H. Jiang, Y. Li, C. Roberts, M. Lu, A. Kapke, and M. Chopp Endothelial Nitric Oxide Synthase Regulates Brain-Derived Neurotrophic Factor Expression and Neurogenesis after Stroke in Mice J. Neurosci., March 2, 2005; 25(9): 2366 - 2375. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. Chan, K. O'Donoghue, J. de la Fuente, I. A. Roberts, S. Kumar, J. E. Morgan, and N. M. Fisk Human Fetal Mesenchymal Stem Cells as Vehicles for Gene Delivery Stem Cells, January 1, 2005; 23(1): 93 - 102. [Abstract] [Full Text] [PDF] |
||||
![]() |
O. Lindvall and Z. Kokaia Recovery and Rehabilitation in Stroke: Stem Cells Stroke, November 1, 2004; 35(11_suppl_1): 2691 - 2694. [Abstract] [Full Text] [PDF] |
||||
![]() |
W.-C. Shyu, S.-Z. Lin, H.-I Yang, Y.-S. Tzeng, C.-Y. Pang, P.-S. Yen, and H. Li Functional Recovery of Stroke Rats Induced by Granulocyte Colony-Stimulating Factor-Stimulated Stem Cells Circulation, September 28, 2004; 110(13): 1847 - 1854. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. F. Ji, B. P. He, S. T. Dheen, and S. S. W. Tay Interactions of Chemokines and Chemokine Receptors Mediate the Migration of Mesenchymal Stem Cells to the Impaired Site in the Brain After Hypoglossal Nerve Injury Stem Cells, May 1, 2004; 22(3): 415 - 427. [Abstract] [Full Text] [PDF] |
||||
![]() |
W.-R. Schabitz, C. Berger, R. Kollmar, M. Seitz, E. Tanay, M. Kiessling, S. Schwab, and C. Sommer Effect of Brain-Derived Neurotrophic Factor Treatment and Forced Arm Use on Functional Motor Recovery After Small Cortical Ischemia Stroke, April 1, 2004; 35(4): 992 - 997. [Abstract] [Full Text] [PDF] |
||||
![]() |
S.-W. Jeong, K. Chu, K.-H. Jung, S. U. Kim, M. Kim, and J.-K. Roh Human Neural Stem Cell Transplantation Promotes Functional Recovery in Rats With Experimental Intracerebral Hemorrhage Stroke, September 1, 2003; 34(9): 2258 - 2263. [Abstract] [Full Text] [PDF] |
||||
![]() |
I. M. Barbash, P. Chouraqui, J. Baron, M. S. Feinberg, S. Etzion, A. Tessone, L. Miller, E. Guetta, D. Zipori, L. H. Kedes, et al. Systemic Delivery of Bone Marrow-Derived Mesenchymal Stem Cells to the Infarcted Myocardium: Feasibility, Cell Migration, and Body Distribution Circulation, August 19, 2003; 108(7): 863 - 868. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Saito, J.-Q. Kuang, C. C. H. Lin, and R. C.-J. Chiu Transcoronary implantation of bone marrow stromal cells ameliorates cardiac function after myocardial infarction J. Thorac. Cardiovasc. Surg., July 1, 2003; 126(1): 114 - 122. [Abstract] [Full Text] [PDF] |
||||
![]() |
L. Zhang, Z. G. Zhang, R. L. Zhang, M. Lu, M. Krams, and M. Chopp Effects of a Selective CD11b/CD18 Antagonist and Recombinant Human Tissue Plasminogen Activator Treatment Alone and in Combination in a Rat Embolic Model of Stroke Stroke, July 1, 2003; 34(7): 1790 - 1795. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Wehner, M. Bontert, I. Eyupoglu, K. Prass, M. Prinz, F. F. Klett, M. Heinze, I. Bechmann, R. Nitsch, F. Kirchhoff, et al. Bone Marrow-Derived Cells Expressing Green Fluorescent Protein under the Control of the Glial Fibrillary Acidic Protein Promoter Do Not Differentiate into Astrocytes In Vitro and In Vivo J. Neurosci., June 15, 2003; 23(12): 5004 - 5011. [Abstract] [Full Text] [PDF] |
||||
![]() |
L. Zhang, Z. G. Zhang, R. Zhang, D. Morris, M. Lu, B. S. Coller, and M. Chopp Adjuvant Treatment With a Glycoprotein IIb/IIIa Receptor Inhibitor Increases the Therapeutic Window for Low-Dose Tissue Plasminogen Activator Administration in a Rat Model of Embolic Stroke Circulation, June 10, 2003; 107(22): 2837 - 2843. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. Chen, Z. G. Zhang, Y. Li, L. Wang, Y. X. Xu, S. C. Gautam, M. Lu, Z. Zhu, and M. Chopp Intravenous Administration of Human Bone Marrow Stromal Cells Induces Angiogenesis in the Ischemic Boundary Zone After Stroke in Rats Circ. Res., April 4, 2003; 92(6): 692 - 699. [Abstract] [Full Text] [PDF] |
||||
![]() |
D. A. Rempe and T. A. Kent Using bone marrow stromal cells for treatment of stroke Neurology, August 27, 2002; 59(4): 486 - 486. [Full Text] [PDF] |
||||
![]() |
Y. Li, J. Chen, X. G. Chen, L. Wang, S. C. Gautam, Y. X. Xu, M. Katakowski, L. J. Zhang, M. Lu, N. Janakiraman, et al. Human marrow stromal cell therapy for stroke in rat: Neurotrophins and functional recovery Neurology, August 27, 2002; 59(4): 514 - 523. [Abstract] [Full Text] [PDF] |
||||
![]() |
D. J. Gladstone, S. E. Black, and A. M. Hakim Toward Wisdom From Failure: Lessons From Neuroprotective Stroke Trials and New Therapeutic Directions Stroke, August 1, 2002; 33(8): 2123 - 2136. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Saito, J.-Q. Kuang, B. Bittira, A. Al-Khaldi, and R. C.-J. Chiu Xenotransplant cardiac chimera: immune tolerance of adult stem cells Ann. Thorac. Surg., July 1, 2002; 74(1): 19 - 24. [Abstract] [Full Text] [PDF] |
||||
![]() |
D. C. Hess, W. D. Hill, A. Martin-Studdard, J. Carroll, J. Brailer, and J. Carothers Bone Marrow as a Source of Endothelial Cells and NeuN-Expressing Cells After Stroke Stroke, May 1, 2002; 33(5): 1362 - 1368. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. Chen, P. R. Sanberg, Y. Li, L. Wang, M. Lu, A. E. Willing, J. Sanchez-Ramos, and M. Chopp Intravenous Administration of Human Umbilical Cord Blood Reduces Behavioral Deficits After Stroke in Rats Stroke, November 1, 2001; 32(11): 2682 - 2688. [Abstract] [Full Text] [PDF] |
||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
Stroke Home | Subscriptions | Archives | Feedback | Authors | Help | AHA Journals Home | Search Copyright © 2001 American Heart Association, Inc. All rights reserved. Unauthorized use prohibited. |