Bone Marrow Stromal Cells Transplantation Research Articles

Timing and cell dose determine therapeutic effects of bone marrow stromal cell transplantation in rat model of cerebral infarct

Stereotactic transplantation of bone marrow stromal cells (BMSCs) enables efficient delivery to the infarct brain. This study was aimed to assess its optimal timing and cell dose for ischemic stroke. The BMSCs were harvested from the green fluorescent protein-transgenic rats and were labeled with quantum dots. The BMSCs (1 × 10(5) or 1 × 10(6) ) were stereotactically transplanted into the ipsilateral striatum of the rats subjected to permanent middle cerebral artery occlusion at 1 or 4 weeks post-ischemia. Motor function was serially assessed. Using in vivo near infrared (NIR) fluorescence imaging, the engrafted BMSCs were visualized at 3 weeks post-transplantation. Immunohistochemistry was performed to evaluate their fate. Functional recovery was significantly enhanced when both low and high doses of BMSCs were transplanted at 1 week post-ischemia, but such therapeutic effects were observed only when the high-dose BMSCs were transplanted at 4 weeks post-ischemia. Both optical imaging and immunohistochemistry revealed their better engraftment in the peri-infarct area when the high-dose BMSCs were transplanted at 1 or 4 weeks post-ischemia. These findings strongly suggest the importance of timing and cell dose to yield therapeutic effects of BMSC transplantation for ischemic stroke. Earlier transplantation requires a smaller number of donor cells for beneficial effects.

Intracerebroventricular transplanted bone marrow stem cells survive and migrate into the brain of rats with Parkinson's disease.

In this study, 6-hydroxydopamine was stereotaxically injected into the right substantia nigra compact and ventral tegmental area of rats to establish Parkinson's disease models. The rats then received a transplantation of bone marrow stromal cells that were previously isolated, cultured and labeled with 5-bromo-2’-deoxyuridine in vitro. Transplantation of the bone marrow stromal cells significantly decreased apomorphine-induced rotation time and the escape latency in the Morris water maze test as compared with rats with untreated Parkinson's disease. Immunohistochemical staining showed that, 5-bromo-2’-deoxyuridine-immunoreactive cells were present in the lateral ventricular wall and the choroid plexus 1 day after transplantation. These immunoreactive cells migrated to the surrounding areas of the lateral cerebral ventricle along the corpus callosum. The results indicated that bone marrow stromal cells could migrate to tissues surround the cerebral ventricle via the cerebrospinal fluid circulation and fuse with cells in the brain, thus altering the phenotype of cells or forming neuron-like cells or astrocytes capable of expressing neuron-specific proteins. Taken together, the present findings indicate that bone marrow stromal cells transplanted intracerebroventricularly could survive, migrate and significantly improve the rotational behavior and cognitive function of rats with experimentally induced Parkinson's disease.

Bone marrow stromal cell transplantation combined with angiotensin-converting enzyme inhibitor treatment in rat with acute myocardial infarction and the role of insulin-like growth factor-1

We investigated bone marrow stromal cell (BMSC) transplantation combined with angiotensin-converting enzyme inhibitor (ACEI) treatment in acute myocardial infarction (AMI) and the role of insulin-like growth factor-1 (IGF-1). AMI models were established in Sprague-Dawley rats by ligation of the left anterior descending coronary artery and grouped into blank control (BC), ACEI treatment (ACEI), BMSC transplantation (BMSC) and BMSC transplantation plus ACEI (combined). Perindopril (2.5 mg/kg) was administered by gavage to ACEI and combined groups from the day after AMI. BMSC (2 × 10(8)) were injected into the border of the MI area a week later in the BMSC and combined groups. After 4 weeks, hemodynamics in the BMSC and combined groups were significantly improved (P < 0.05 versus BC), with the greatest improvement in the combined group (P < 0.05). In addition, an increased number of BMSC survived in the combined group (P < 0.05 versus BMSC). A proportion of BMSC was positive for troponin T, as detected by immunofluorescence. The number of apoptotic cardiomyocytes was decreased in the BMSC and ACEI groups, and even further in the combined group (P < 0.05). IGF-1 expression was up-regulated in the BMSC and combined groups (P < 0.05 versus BC), but not in the ACEI group. B cell lymphoma-2 (Bcl-2) expression was up-regulated in the ACEI, BMSC and combined groups, with the highest expression in the combined group (P < 0.05). Our results show that BMSC engrafted in AMI can survive well and secrete IGF-1 and preserve cardiac function significantly. These data suggest that BMSC transplantation inhibits apoptosis of cardiomyocytes by up-regulation of Bcl-2 expression in the myocardium, and this effect might be sensitized by ACEI.

The severity of brain damage determines bone marrow stromal cell therapy efficacy in a traumatic brain injury model

Patients who survive traumatic brain injury (TBI) can undergo serious sensorial and motor function deficits. Once damage occurs, there is no effective treatment to bring patients to full recovery. Recent studies, however, show bone marrow stromal cells (BMSC) as a potential therapy for TBI. This study was designed to determine whether the degree of neurologic deficits influences the efficacy of cell therapy using intracerebral transplantation of BMSC in an experimental model of chronically established TBI. Adult Wistar rats were subjected to weight-drop impact causing TBI. Two months later, the animals were classified according to levels of neurologic deficits. To achieve this, we used two different functional tests: the modified Neurologic Severity Score test and internal zone Permanence Time in Video-Tracking-Box analysis. Saline only or saline containing BMSC was injected into injured brain tissue of the animals that were classified having moderate or severe neurologic damage depending on the level of established functional deficits. All experimental groups were evaluated in the course of the following 2 months to study the efficacy of BMSC administration. The animals were then killed and their brains were studied. Our results showed that significant functional improvement was seen when BMSC was injected into animals with moderate brain damage, but no significant improvement was found in animals with severe functional deficits when compared with controls. These findings suggest that the severity of neurologic damage may determine the potential effect of cell therapy when applied to chronically established TBI.

Safety of Autologous Bone Marrow Stromal Cell Transplantation in Dogs with Acute Spinal Cord Injury

To assess the feasibility and safety of transplantation of autologous bone marrow stromal cell (BMSC) in dogs with acute spinal cord injury (SCI). An open-label single-arm trial. Dogs (n = 7) with severe SCI from T6 to L5, caused by vertebral fracture and luxation. Decompressive and stabilization surgery was performed on dogs with severe SCI caused by vertebral fracture and luxation. Autologous BMSCs were obtained from each dog's femur, cultured, and then injected into the lesion in the acute stage. Adverse events and motor and sensory function were observed for >1 year after SCI. Follow-up was 29-62 months after SCI. No complications (eg, infection, neuropathic pain, worsening of neurologic function) were observed. Two dogs walked without support, but none of the 7 dogs had any change in sensory function. Autologous BMSC transplantation is feasible and safe in dogs with acute SCI. Further studies are needed to determine the efficacy of this therapy.

Functional recovery and neuronal regeneration of a rat model of epilepsy by transplantation of Hes1-down regulated bone marrow stromal cells

Gamma-amino butyric acid (GABA)ergic cells play an important inhibitory role in epilepsy. Until now, there are no reports on promoting transplanted bone marrow stromal cells (BMSCs) to differentiate into GABAergic cells for treatment of epilepsy. In this study, hairy and enhancer of split 1 (Hes1)-down regulated BMSCs (H-BMSCs) were transplanted into an epileptic rat model to induce GABAergic cells differentiation to improve the function recovery and neuronal regeneration. First, Hes1 expression in isolated BMSCs was down regulated by Hes1 siRNA. Then, the H-BMSCs were labeled with 5-bromo-2′-deoxyuridine (BrdU) and transplanted into the lateral ventricle of pilocarpine-induced epileptic rats. To evaluate the therapeutic effects, behavior and electroencephalography (EEG) of the recipient rats were monitored in the following 4weeks, followed by histological confirmation. The results showed that the rate of mortality, frequency of spontaneous recurrent seizures (SRS) and incidence of epileptiform waves presented a tendency to decrease after H-BMSCs transplantation. The histology results showed that (1) the transplanted H-BMSCs which migrated to the adjacent parahippocampal cortical areas expressed glutamate decarboxylase (GAD) 67, neuron-specific enolase (NSE) and some glial fibrillary acidic protein (GFAP), and (2) the neuronal density of corresponding cortical areas was significantly increased (P<0.01 VS. experimental group I or positive control group). Given these results and other advantages of BMSCs, such as easy harvest and minimal immunogenicity, transplantation of H-BMSCs could be a promising approach to improve the functional recovery and neuronal regeneration of epileptic model in the early stage.

Transplanting bone mesenchymal stem cells via posterior intercostal arteries in a rabbit model of spinal injury

Objective To evaluate the feasibility of transplantation of bone marrow stromal cells (BMSCs) via posterior intercostal arteries for the treatment of spinal cord injury (SCI). Methods BMSCs which had been obtained from 2 rabbits were expanded after intermediate culture.All BMSCs were labeled with Feridex and transfected by green fluorescent protein (GFP).Another 20 rabbits of SCI models were randomized into 2 groups:a BMSCs group in which BMSCs were injected into the posterior intercostal arteries and a control group in which no treatment was adopted.The injected BMSCs were monitored by MRI in vivo.Fluorescence images and electron micrographs of the spinal cord were taken to confirm the MRI results and nerve conduction of the damaged spinal cord was asessed by nerve electrophysiological examination.On 7 and 14 days post-experiment,the latency and amplitude of motion evoked potential (MEP) in 2 groups was measured. Results MRI showed migration of BMSCs to the injury sites 14 days post-transplantation,which was confirmed by fluorescent microscpy.The myelin regenerated by oligodendrocytes was shown containing Fetidex particles as observed by electron microscopy.There was no significant difference between the 2 groups on day 7 in nerve conduction of the damaged spinal cord ( P ＞ 0.05).On the 14 day,however,the MEP latency and amplitude in the control group and the experiment group were 26.7 ± 0.43 vs.17.9 ± 0.46 ms ( n =10) and 39.9 ± 0.65 vs.43.3 ± 0.67μv ( n =10) respectively,with significant differences between the 2 groups (P ＜0.05). Conclusion Transplanting BMSCs via postetior intercostal arteries is a noninvasive and efficient intervention for treatment of SCI,available for clinical experiment. Key words: Spinal cord injuries; Rabbits; Bone marrow cells

Effect of bone marrow stromal cells transplantation on sensorimotor and autonomic function in complete spinal cord transection injury rats

International Journal of Developmental NeuroscienceVolume 30, Issue 8 p. 659-659 Abstract Effect of bone marrow stromal cells transplantation on sensorimotor and autonomic function in complete spinal cord transection injury rats First published: 31 March 2012 https://doi.org/10.1016/j.ijdevneu.2012.03.293AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume30, Issue8Abstracts to the 19th Biennial Meeting of the International Society for Developmental Neuroscience, 11-14 January 2012, Mumbai, IndiaDecember 2012Pages 659-659 RelatedInformation

Effect of free NgR-modified bone marrow stromal cell transplantation on axon regeneration after spinal cord injury

Objective To evaluate the effect of free NgR-modified bone marrow stromal cell (BMSC) transplantation on axon regeneration in rats after spinal cord injury. Methods Genes encoding free NgR protein were cloned and transduced into BMSCs at passage 3 using a lentivirus vector.Indirect immunofluorescence was used to detect the expression of free NgR protein.Meanwhile a spinal cord contusion model was established in 36 adult Sprague-Dawley rats at the T10 segment.The rats were then divided randomly into an experimental group and a control group.NgR + BMSCs were transplanted into the injured site 1 week post-trauma in the experimental group.BMSCs were also transplanted at the same time into the control group.Expression of free NgR at the injury site was detected by immunohistochemical staining at 1 week post-transplantation.The functional recovery of both groups was evaluated at 4 and 6 weeks post-transplantation.Longitudinal sections of the spinal cord were studied for axon regeneration using horseradish peroxidase staining. Results Expression of free NgR was found in the cell plasma of BMSCs by indirect immunofluorescence post-transfection.Positive immunohistochemical staining for NgR was found at the transplant site in the experimental group 1 week post-transplantation.Better axon plasticity could be observed in the experimental group.The Basso-Beattie-Bresnahan scoring of the experimental group was significantly higher than that of the controls at both observation times. Conclusions Free NgR-modified BMSCs can prompt injured axons to regenerate and thus to promote the recovery of neurological function.This might provide a new strategy to treat spinal cord injury. Key words: NgR protein; Bone marrow stromal cells; Axon regeneration; Spinal cord injury; Rehabilitation

Icariin recovers the osteogenic differentiation and bone formation of bone marrow stromal cells from a rat model of estrogen deficiency-induced osteoporosis.

A number of recent studies have suggested that icariin (ICA), a class of phytochemical with numerous biological activities, may exert protective effects against postmenopausal bone loss. However, it remains unclear whether ICA regulates or improves the osteoblastic function of bone marrow stromal cells (BMSCs) in the treatment and prevention of osteoporosis. In the present study, the osteogenic differentiation of BMSCs from ovariectomy (OVX) rats was found to be significantly decreased in vitro compared with that in rats that had undergone a sham operation. Treatment with ICA at a dose of 10-5 M was shown to restore the osteogenic differentiation of BMSCs in OVX rats. The results indicated that ICA restored the differentiation and mineralization capacity of OVX-BMSCs, which had been induced by estrogen deficiency. The effects of this compound on alkaline phosphatase (ALP) activity and calcium deposition were also measured at various time points. The number of colonies and areas that stained positive for ALP expression, and mineralized bone nodules were analyzed histochemically at 14 and 21 days after the osteogenic induction. The expression of the runt-related transcription factor 2 and osterix bone metabolism biomarker proteins and genes were detected by western blotting and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The expression of factors involved in the estrogen signaling pathway, estrogen receptor α (ERα), progesterone receptor (PR) and trefoil factor 1 (PS-2), was also detected by western blotting and RT-qPCR. ICA enhanced the expression of ERα, PR, PS-2 in OVX‑BMSCs, but this effect was abrogated when ICI 182780, an ER antagonist was added. Transplantation of BMSCs into nude mice demonstrated that ICA restored the osteogenic capability of OVX‑BMSCs in vivo. Therefore, it may be that ICA acts through the estrogen pathway in order to improve and restore the osteogenic differentiation and mineralization of OVX‑BMSCs, which are inhibited by estrogen deficiency and increasing age.

An In Vivo Characterization of Trophic Factor Production Following Neural Precursor Cell or Bone Marrow Stromal Cell Transplantation for Spinal Cord Injury

Cellular transplantation strategies for repairing the injured spinal cord have shown consistent benefit in preclinical models, and human clinical trials have begun. Interactions between transplanted cells and host tissue remain poorly understood. Trophic factor secretion is postulated a primary or supplementary mechanism of action for many transplanted cells, however, there is little direct evidence to support trophin production by transplanted cells in situ. In the present study, trophic factor expression was characterized in uninjured, injured-untreated, injured-treated with transplanted cells, and corresponding control tissue from the adult rat spinal cord. Candidate trophic factors were identified in a literature search, and primers were designed for these genes. We examined in vivo trophin expression in 3 paradigms involving transplantation of either brain or spinal cord-derived neural precursor cells (NPCs) or bone marrow stromal cells (BMSCs). Injury without further treatment led to a significant elevation of nerve growth factor (NGF), leukemia inhibitory factor (LIF), insulin-like growth factor-1 (IGF-1), and transforming growth factor-β1 (TGF-β1), and lower expression of vascular endothelial growth factor isoform A (VEGF-A) and platelet-derived growth factor-A (PDGF-A). Transplantation of NPCs led to modest changes in trophin expression, and the co-administration of intrathecal trophins resulted in significant elevation of the neurotrophins, glial-derived neurotrophic factor (GDNF), LIF, and basic fibroblast growth factor (bFGF). BMSCs transplantation upregulated NGF, LIF, and IGF-1. NPCs isolated after transplantation into the injured spinal cord expressed the neurotrophins, ciliary neurotrophic factor (CNTF), epidermal growth factor (EGF), and bFGF at higher levels than host cord. These data show that trophin expression in the spinal cord is influenced by injury and cell transplantation, particularly when combined with intrathecal trophin infusion. Trophins may contribute to the benefits associated with cell-based repair strategies for spinal cord injury.

Abstract 119: Bone Marrow Stromal Cell Transplantation Enhances Recovery of Local Glucose Metabolism after Cerebral Infarct in Rats: A 18 F-FDG PET Study

Background: Recent studies have indicated that bone marrow stromal cells (BMSC) have the potential to improve neurologic function when transplanted into animal models of central nervous system disorders. However, no methods have been established to objectively evaluate the beneficial effect of BMSC transplantation on the host brain in clinical situation. Therefore, this study was aimed to assess whether the BMSC significantly improve local glucose metabolism in the peri-infarct area on 18 F-fluorodeoxyglucose (FDG) PET, when transplanted into the infarct brain of rats. Methods: The BMSC were harvested from green fluorescent protein (GFP)-transgenic rats and were cultured. The rats were subjected to permanent middle cerebral artery (MCA) occlusion. The BMSC or vehicle was transplanted into the ipsilateral striatum at 7 days after the onset of MCA occlusion. Using 18 F-FDG PET, local glucose metabolism was semi-quantitatively and serially measured at 6 days post-ischemia and at 4 weeks after BMSC transplantation. Motor function was serially evaluated throughout the experiments. The distribution and fate of engrafted cells were examined, using fluorescence immunohistochemistry at 4 weeks after BMSC transplantation. Simultaneously, the expression of brain-type glucose transporter (GLUT1 and GLUT3) was also evaluated. Results: Local glucose metabolism was significantly lower in the dorsal peri-infarct neocortex at 6 days post-ischemia. However, BMSC transplantation significantly accelerated the recovery of local glucose metabolism in that area at 4 weeks after transplantation. Thus, the ratio of glucose metabolism in the ipsilateral to contralateral dorsal neocortex significantly increased from 72.5 ± 4.2% to 78.7 ± 4.0% in the vehicle group (P&lt;0.01). On the other hands, the values more pronouncedly increased from 72.7 ± 4.4% to 87.7 ± 5.3% in the BMSC group (P&lt;0.01). The BMSC were engrafted in the boundary zone of cerebral infarct and also expressed the phenotypes of neural cells. BMSC transplantation significantly prevented the overexpression of GLUT1 in the endothelium and of GLUT3 in the neurons in that area. Conclusion: These findings strongly suggest that BMSC may enhance the recovery of local glucose metabolism in the peri-infarct area when directly transplanted into the infarct brain at clinically relevant timing. The recovery of local glucose metabolism may inhibit pathological upregulation of GLUT1 and GLUT3. 18 F-FDG PET may be the candidate of valuable modalities to scientifically prove the beneficial effects of BMSC transplantation on the host brain in clinical situation.

Therapeutic Effects of Intra-Arterial Delivery of Bone Marrow Stromal Cells in Traumatic Brain Injury of Rats—In Vivo Cell Tracking Study by Near-Infrared Fluorescence Imaging

A noninvasive and effective route of cell delivery should be established to yield maximal therapeutic effects for central nervous system (CNS) disorders. To elucidate whether intra-arterial delivery of bone marrow stromal cells (BMSCs) significantly promotes functional recovery in traumatic brain injury (TBI) in rats. Rat BMSCs were transplanted through the ipsilateral internal carotid artery 7 days after the onset of cortical freezing injury. The BMSCs were labeled with fluorescent dye, and in vivo optical imaging was employed to monitor the behaviors of cells for 4 weeks after transplantation. Motor function was assessed for 4 weeks, and the transplanted BMSCs were examined using immunohistochemistry. In vivo optical imaging and histologic analysis clearly demonstrated that the intra-arterially injected BMSCs were engrafted during the first pass without systemic circulation, and the transplanted BMSCs started to migrate from the cerebral capillary bed to the injured CNS tissue within 3 hours. Intra-arterial BMSC transplantation significantly promoted functional recovery after cortical freezing injury. A subgroup of BMSCs expressed the phenotypes of neurons, astrocytes, and endothelial cells around the injured neocortex 4 weeks after transplantation. Intra-arterial transplantation may be a valuable option for prompt, noninvasive delivery of BMSCs to the injured CNS tissue, enhancing functional recovery after TBI. In vivo optical imaging may provide important information on the intracerebral behaviors of donor cells by noninvasive, serial visualization.

Transplanted bone marrow stromal cells protect neurovascular units and ameliorate brain damage in stroke‐prone spontaneously hypertensive rats

This study was aimed to assess whether bone marrow stromal cells (BMSC) could ameliorate brain damage when transplanted into the brain of stroke-prone spontaneously hypertensive rats (SHR-SP). The BMSC or vehicle was stereotactically engrafted into the striatum of male SHR-SP at 8 weeks of age. Daily loading with 0.5% NaCl-containing water was started from 9 weeks. MRIs and histological analysis were performed at 11 and 12 weeks, respectively. Wistar-Kyoto rats were employed as the control. As a result, T2-weighted images demonstrated neither cerebral infarct nor intracerebral hemorrhage, but identified abnormal dilatation of the lateral ventricles in SHR-SP. HE staining demonstrated selective neuronal injury in their neocortices. Double fluorescence immunohistochemistry revealed that they had a decreased density of the collagen IV-positive microvessels and a decreased number of the microvessels with normal integrity between basement membrane and astrocyte end-feet. BMSC transplantation significantly ameliorated the ventricular dilatation and the breakdown of neurovascular integrity. These findings strongly suggest that long-lasting hypertension may primarily damage neurovascular integrity and neurons, leading to tissue atrophy and ventricular dilatation prior to the occurrence of cerebral stroke. The BMSC may ameliorate these damaging processes when directly transplanted into the brain, opening the possibility of prophylactic medicine to prevent microvascular and parenchymal-damaging processes in hypertensive patients at higher risk for cerebral stroke.

Therapeutic Potential of Canine Bone Marrow Stromal Cells (BMSCs) in the Carbon Tetrachloride (CCl&lt;sub&gt;4&lt;/sub&gt;) Induced Chronic Liver Dysfunction Mouse Model

Regenerative medicine using bone marrow cells is an attractive therapy for the cure of patients with severe liver disease. Here, we show the therapeutic potential of canine bone marrow stromal cells (BMSCs) in mouse models of CCl(4)-induced chronic liver dysfunction. We used two different models for xenotransplantation, nude mice and cyclosporine A (CSA) immunosuppressed mice. Serum parameters from a standard liver panel were not improved following transplantation. However, fibrotic liver lesions with severe inflammation were decreased in CCl(4)-treated CSA mice following BMSC transplantation. Effective migration of transplanted canine BMSCs was limited to persistently injured liver in CCl(4)-treated CSA mice, where they may be effective in resolving inflammatory fibrotic lesions. These results suggest that canine BMSCs are an effective cell source for liver regeneration.

Expression of DeltaNp73 in hippocampus of APP/PS1 transgenic mice following GFP-BMSCs transplantation

Objective: To study the effect of hippocampal bone marrow stromal cells (GFP-BMSCs) transplantation on spatial memory and DeltaNp73 expression in APP/PS1 transgenic mice.Methods: Twelve APP/PS1 transgenic mice randomly received either 10 μl GFP-BMSCs suspension in medium (GFP-BMSCs transplantation group) or 10 μl complete medium (sham-operated group). Learning and memory function of mice in both groups were observed and tested in Morris water maze experiment at 2 weeks after surgery. Senile plaques and DeltaNp73 protein in hippocampuses were determined by immunohistochemistry and western blot at 3 weeks after surgery, respectively.Results: APP/PS1 mice treated with BMSCs performed significantly better on the water maze test than those in sham-operated group (P<0·05). Immunohistochemistry showed that GFP-BMSCs distributed uniformly and the number of Alzheimer’s senile plaques reduced after transplantation. Western blot showed that quantified DeltaNp73 protein expression was significantly higher in BMSCs transplantation group when compared with sham-operated group (P<0·01).Conclusions: Our results suggest that BMSCs transplatation could retard Alzheimer’s disease (AD) like pathology and upregulate DeltaNp73 expression in hippocampuses of APP/PS1 transgenic mice. GFP-BMSCs transplantation will be a potential treatment for AD.

Intravenously transplanted bone marrow stromal cells promote recovery of lower urinary tract function in rats with complete spinal cord injury

There is increasing evidence that intravenously injected neural progenitor cells promote recovery of bladder function in rodents, following contusive spinal cord injury through migrating into the injured spinal cord tissue and differentiating into central nervous system cells. The present study was aimed to clarify whether intravenously transplanted bone marrow stromal cells (BMSCs) could improve lower urinary tract (LUT) function in rats with spinal cord transection (SCT). A total of 22 rats underwent experimentation in three groups, including group 1-sham operation, group 2 (BMSC)-SCT plus BrdU (5-bromo-2'-deoxyuridine) labeled BMSCs transplantation at day 9 after SCT, group 3-SCT control. All rats were investigated urodynamically on day 28 after transplantation. BMSCs identified by BrdU immunohistochemistry survived in the injured spinal cord and lumbar level 3-4 (L(3-4)). Voiding pressure, episodes of non-voiding contractions and residual urine volumes were significantly decreased in BMSC rats, compared with the controls. Bladder capacity was similar in both groups. In four out of eight BMSC rats and one out of seven controls, the tonic and bursting external urethral sphincter electromyographic activity were detected during cystometry. Silent periods during bursting were shorter and activity periods were longer in BMSC rats compared with sham rats. Intravenously transplanted BMSCs survived in the L(3-4) and had beneficial effects on the recovery of LUT function in the rats after SCT.

Isolation and characterization of neural crest-derived stem cells from dental pulp of neonatal mice.

Dental pulp stem cells (DPSCs) are shown to reside within the tooth and play an important role in dentin regeneration. DPSCs were first isolated and characterized from human teeth and most studies have focused on using this adult stem cell for clinical applications. However, mouse DPSCs have not been well characterized and their origin(s) have not yet been elucidated. Herein we examined if murine DPSCs are neural crest derived and determined their in vitro and in vivo capacity. DPSCs from neonatal murine tooth pulp expressed embryonic stem cell and neural crest related genes, but lacked expression of mesodermal genes. Cells isolated from the Wnt1-Cre/R26R-LacZ model, a reporter of neural crest-derived tissues, indicated that DPSCs were Wnt1-marked and therefore of neural crest origin. Clonal DPSCs showed multi-differentiation in neural crest lineage for odontoblasts, chondrocytes, adipocytes, neurons, and smooth muscles. Following in vivo subcutaneous transplantation with hydroxyapatite/tricalcium phosphate, based on tissue/cell morphology and specific antibody staining, the clones differentiated into odontoblast-like cells and produced dentin-like structure. Conversely, bone marrow stromal cells (BMSCs) gave rise to osteoblast-like cells and generated bone-like structure. Interestingly, the capillary distribution in the DPSC transplants showed close proximity to odontoblasts whereas in the BMSC transplants bone condensations were distant to capillaries resembling dentinogenesis in the former vs. osteogenesis in the latter. Thus we demonstrate the existence of neural crest-derived DPSCs with differentiation capacity into cranial mesenchymal tissues and other neural crest-derived tissues. In turn, DPSCs hold promise as a source for regenerating cranial mesenchyme and other neural crest derived tissues.

In vivo formation of bone and haematopoietic territories by transplanted human bone marrow stromal cells generated in medium with and without osteogenic supplements.

Autologous transplantation of human bone marrow stromal cells (BMSCs) has been successfully used for bone reconstruction. However, in order to advance this approach into the mainstream of bone tissue engineering, the conditions for BMSC cultivation and transplantation must be optimized. In a recent report, cultivation with dexamethasone (Dex) significantly increased bone formation by human BMSCs in vivo. Based on this important conclusion, we analysed the data accumulated by our laboratory, where human BMSCs have been routinely generated using media both with and without a combination of two osteogenic supplements: Dex at 10(-8) m and ascorbic acid phosphate (AscP) at 10(-4) m. Our data demonstrate that for 22/24 donors, BMSC strains propagated with and without Dex/AscP formed similar amounts of bone in vivo. Thus, human BMSCs do not appear to need to be induced to osteogenic differentiation ex vivo prior to transplantation. Similarly, for 12/14 donors, BMSC strains cultured with and without Dex/AscP formed haematopoietic territories to a comparable extent. While Dex/AscP did not increase bone formation, they significantly stimulated BMSC in vitro proliferation without affecting the number of BMSC colonies formed by the colony-forming units-fibroblasts. We conclude that for the substantial majority of donors, Dex/AscP have no effect on the ability of BMSCs to form bone and myelosupportive stroma in vivo. However, due to increased BMSC proliferation, the total osteogenic population obtained from a single marrow sample is larger after cultivation with Dex/AscP than without them. Secondary to increased BMSC proliferation, Dex/AscP may stimulate bone formation if BMSCs and/or the transplantation system are less than optimal. Published 2011. This article is a U.S. Government work and is in the public domain in the USA.

Intracerebral, but not intravenous, transplantation of bone marrow stromal cells enhances functional recovery in rat cerebral infarct: An optical imaging study

Recent studies have indicated that bone marrow stromal cells (BMSC) may improve neurological function when transplanted into an animal model of CNS disorders, including cerebral infarct. However, there are few studies that evaluate the therapeutic benefits of intracerebral and intravenous BMSC transplantation for cerebral infarct. This study was aimed to clarify the favorable route of cell delivery for cerebral infarct in rats. The rats were subjected to permanent middle cerebral artery occlusion. The BMSC were labeled with near infrared (NIR)-emitting quantum dots and were transplanted stereotactically (1 × 10⁶ cells) or intravenously (3 × 10⁶ cells) at 7 days after the insult. Using in vivo NIR fluorescence imaging technique, the behaviors of BMSC were serially visualized during 4 weeks after transplantation. Motor function was also assessed. Immunohistochemistry was performed to evaluate the fate of the engrafted BMSC. Intracerebral, but not intravenous, transplantation of BMSC significantly enhanced functional recovery. In vivo NIR fluorescence imaging could clearly visualize their migration toward the cerebral infarct during 4 weeks after transplantation in the intracerebral group, but not in the intravenous, group. The BMSC were widely distributed in the ischemic brain and some of them expressed neural cell markers in the intracerebral group, but not in the intravenous group. These findings strongly suggest that intravenous administration of BMSC has limited effectiveness at clinically relevant timing and intracerebral administration should be chosen for patients with ischemic stroke, although further studies would be warranted to establish the treatment protocol.