hUC-MSCs Transplantation Research Articles

A combination of GDNF and hUCMSC transplantation loaded on SF/AGs composite scaffolds for spinal cord injury repair

Spinal cord injury (SCI) is a severe trauma for which no effective treatment is currently available. In this study, a composited treatment system was prepared using a silk fibroin/alginates/glial cell line-derived neurotrophic factor (SF/AGs/GDNF) scaffold seeded with human umbilical cord mesenchymal stem cells (hUCMSCs) and the combined therapeutic effects of the composite scaffold to repair SCI rats were evaluated. The use of SF as a scaffold material could act as a biomimetic platform allowing neurons to properly accommodate and rebuild the target tissue. The SF/AGs/GDNF scaffold had the best sustained-release function and the AGs were the key determining factor in the controlled release of GDNF. After 8weeks of treatment, the hUCMSCs on SF/AGs/GDNF composite scaffolds could significantly enhance the scar expansion of spinal cord tissue and increased the number of surviving neurons. The combination of GDNF and hUCMSCs transplantation loaded on SF/AGs composite scaffolds exhibited better therapeutic and repair effects to the SCI of rats, compared with the SF/AGs group or GDNF alone on SF/AGs scaffolds. The composite scaffold, GDNF and stem cells could build a bioactive material to form the micro-environment of growth and repair of the neurons. These results may provide a theoretical basis and beneficial exploration for clinical treatment of SCI.

The influence of human umbilical cord mesenchymal stem cells transplantation into vitreous cavity of diabetic rats on retinal morphology and the expression level of glial fibrillary acidic protein and rhodopsin

Objective To observe the influence of human umbilical cord mesenchymal stem cells (hUCMSC) transplantation into vitreous cavity of diabetic rats on the retinal morphology, and the expression of glial fibrillary acidic protein (GFAP) and rhodopsin (RHO). Methods 78 male Sprague-Dawley rats were used. 70 rats were injected with streptozotocin by tail vein injection at a dose of 40 mg/kg to establish the diabetes mellitus model, and another 8 rats were injected with 0.1 mol/L pH 4.0 citric acid buffer at the same dose as the normal control group. After 6 weeks of modeling, 10 rats were taken as the control group of diabetic model. hUCMSC suspension was injected into the right eye vitreous cavity of the remaining 60 rats, and the same volume of Dulbecco′s modified Eagle/F12 medium was injected into the left vitreous cavity as control eyes. 1, 2 and 4 weeks after transplantation, follow-up experiments were performed. The experimental eyes were labeled as U1, U2, and U4 groups, while the control eyes were recorded as D1, D2, D4, and each group consisted of 20 eyes. After paraffin section and hematoxylin-eosin staining, the structure of the retina was observed by optical microscopy and the thickness of the outer nuclear layer and the inner nuclear layer (INL) were measured. The distribution and migration of hUCMSC in rat retina were observed by frozen section-tissue immunofluorescence assay. The mRNA and protein expression of GFAP and RHO in the retina were detected by real-time quantitative polymerase chain reaction (PCR) and Western blot assays. Results The results of optical microscope observation showed the normal structure of retina in normal control group. The retinal nerve fiber layer (NFL) was thinned and the number of retinal ganglion cells (RGC) in the control group of diabetic rats was decreased. The decreased number and disorder arrangement of RGC were observed as well in U1, D1 rats. The RGC number of U2, U4, D2, D4 rats was gradually decreased. Compared with D4 group, the thickness of INL in U4 group was significantly increased (P<0.05). Tissue immunofluorescence assay showed that hUCMSC were distributed along the inner limiting membrane in the retina of the U1 group, while the number of hUCMSC in the U2 group was gradually decreased, mainly in the NFL and ganglion cell layers. Real-time PCR and Western blot data indicated that the relative expression of GFAP mRNA and protein in the diabetic retina was significantly increased, and the relative expression of RHO mRNA and protein decreased gradually in the diabetic model group and the D1, D2, D4 groups. Compared with D2 and D4 groups, the mRNA and protein expression of GFAP in U2 and U4 groups were decreased, and the relative expression of RHO mRNA and protein were all increased (P<0.01). Conclusion hUCMSC could migrate and integrate into the retina, after the transplantation into the vitreous cavity of diabetic rats, which reduced the expression of GFAP, but enhanced the expression of RHO. Key words: Diabetic retinopathy/therapy; Mesenchymal stem cell transplantation; Rhodopsin; Animal experimentation

Human umbilical cord-derived mesenchymal stem cells in acute liver injury: Hepatoprotective efficacy, subchronic toxicity, tumorigenicity, and biodistribution

Umbilical cord-derived mesenchymal stem cells (UC-MSCs) therapy might be an alternative to liver transplantation for acute or chronic liver injury. The aim of this study was to evaluate the efficacy of human UC-MSCs on carbon tetrachloride (CCl4)-induced acute liver injury. In addition, its toxicity, tumorigenicity, and biodistribution were determined. Significant hepatoprotective effects of hUC-MSCs with decreased levels of hepatocellular necrosis and lobular neutrophilic infiltration were found. Regarding the safety of hUC-MSCs, no serious hUC-MSCs-related changes (body weight, food/water consumption, clinical symptom, urinalysis, hematology, clinical chemistry, organ weight, and histopathology) were observed in a 13-week subchronic toxicity study. In a 26-week tumorigenicity study, no mice developed tumor related to hUC-MSCs transplantation up to 1 × 108 cells/kg. In particular, human mitochondrial sequence detection revealed that most hUC-MSCs were cleared from the major organs of the mice at 13 weeks after transplantation. There was no systemic toxicity or neoplastic finding either. Taken together, these results suggested that hUC-MSCs have great potential for future clinical treatment of acute liver disease.

Study of differentiated human umbilical cord-derived mesenchymal stem cells transplantation on rat model of advanced parkinsonism.

The aim of this study was to explore the curative effect of differentiated human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) transplantation on rat of advanced Parkinson disease (PD) model. Human umbilical cord-derived mesenchymal stem cells were cultured and induced differentiation in vitro. The PD rats were established and allocated randomly into 2 groups: differentiated hUC-MSCs groups and physiological saline groups (the control group). Rotation test and immunofluorescence double staining were done. The result showed that hUC-MSCs could differentiate into mature dopamine neurons. Frequency of rotation was significantly less in differentiated hUC-MSCs groups than in normal saline group. After we transplanted these cells into the unilateral lesioned substantia nigra induced by striatal injection of 6-hydroxydopamine and performed in the medial forebrain bundle and ventral tegmental area, nigral tyrosine hydroxylase-positive cells were observed and survival of at least 2months. In addition, transplantation of hUC-MSCs could make an obviously therapeutic effect on PD rats.

Safety of intrathecal transplantation of human umbilical cord mesenchymal stem cells for treating hereditary cerebellar ataxia: study protocol for a randomized controlled trial

Background: Intrathecal transplantation of human umbilical cord mesenchymal stem cells (hUCMSCs) has been shown to be effective for treating spinocerebellar ataxia and can improve patient's neurological function to a certain degree without safety concerns. However, further validation by randomized controlled trials is needed. The present study aims to fill this gap.Methods/Design: A single-center, parallel group, open-label, randomized controlled trial will be performed at General Hospital of Chinese Armed Police Forces, China. A total of 100 patients with hereditary cerebellar ataxia will undergo hUCMSC transplantation in the subarachnoid space through lumbar puncture (hUCMSCs transplantation group) or not (control group). The outcome observation time will be designated within 1 week before transplantation, as well as 1, 3, 6, and 12 months after transplantation. Primary outcomes include neurological function evaluation (International Cooperative Ataxia Rating Scale, Modified Falls Efficacy Scale, Berg Balance Assessment, Tremor Rating Scale, and the Timed Up and Go test) and blood tests. Secondary outcomes include electrophysiology examination, mental state evaluation, and urine testing.Discussion: This trial protocol is designed to provide reference data for the safety and rational use of hUCMSC transplantation in the treatment of hereditary cerebellar ataxia.Trial registration: ClinicalTrials.gov identifier: NCT01489267; registered on 30 October 2011. The study protocol has been approved by the ethics committee of the General Hospital of Chinese Armed Police Forces, China (approval No. 201117).

The protective effects of human umbilical cord mesenchymal stem cells on damaged ovarian function: A comparative study

Numerous studies have reported that human umbilical cord mesenchymal stem cell (hUCMSC) therapy can rescue the structure and function of injured tissues. The aims of this study were to explore the protective role of hUCMSC transplantation in a model of accelerated ovarian aging and to compare 2 methods of transplanting hUCMSCs, i.e. i) via intravenous injection (IV) and ii) in situ ovarian micro injection (MI). Female mice were subjected to superovulation and ozone inhalation to create a model of accelerated ovarian aging with a decline in both the quantity and quality of oocytes. Cells were transplanted via IV or MI, and ovaries were removed after 2 weeks or 1 month of treatment. Ovarian reserve and function were evaluated based on the follicle counts, hormone levels, the estrous cycle, and reproductive performance. Cell tracking, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), real-time polymerase chain reaction (PCR), and Western blot analysis were used to assess the inner mechanisms of injury and repair. Results indicated that ovarian function increased significantly after treatment with hUCMSCs. Immunofluorescence revealed reduced TUNEL staining and a decreased percentage of apoptotic cells. A higher level of expression of anti-apoptotic and antioxidant enzymes was noted in the ovaries of groups treated with hUCMSCs. These parameters were enhanced more when mice were treated with hUCMSCs for 1 month than when they treated with hUCMSCs for 2 weeks. IV was better able to restore ovarian function than MI. These results suggest that both methods of transplantation may improve ovarian function and that IV transplantation of hUCMSCs can significantly improve ovarian function and structural parameters more than MI transplantation of hUCMSCs can.

Effect of Microenvironment on Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Hepatocytes In Vitro and In Vivo.

Human umbilical cord-derived mesenchymal stem cells (hUCMSCs) are considered to be an ideal cell source for cell therapy of many diseases. The aim of this study was to investigate the contribution of the microenvironment to the hepatic differentiation potential of hUCMSCs in vitro and in vivo and to explore their therapeutic use in acute liver injury in rats. We established a new model to simulate the liver tissue microenvironment in vivo using liver homogenate supernatant (LHS) in vitro. This induced environment could drive hUCMSCs to differentiate into hepatocyte-like cells within 7 days. The differentiated cells expressed hepatocyte-specific markers and demonstrated hepatocellular functions. We also injected hUCMSCs into rats with CCl4-induced acute hepatic injury. The hUCMSCs were detected in the livers of recipient rats and expressed the human hepatocyte-specific markers, suggesting that hUCMSCs could differentiate into hepatocyte-like cells in vivo in the liver tissue microenvironment. Levels of biochemistry markers improved significantly after transplantation of hUCMSCs compared with the nontransplantation group (P < 0.05). In conclusion, this study demonstrated that the liver tissue microenvironment may contribute to the differentiation of hUCMSCs into hepatocytes both in vitro and in vivo.

Endogenous antioxidant level of stem cell is important for the transplantation efficacy

Stem cell transplantation has been applied to clinical trials and obtained certain curative effect, but due to the severe environmental such as oxidative stress, the efficiency of stem cell transplantation is still low. However, oxidative stress on stem cells, the influence of the transplantation efficiency and its molecular mechanism are not fully understood. In our recent research, we have confirmed that oxidative stress is the mainly reason that caused the low efficiency of hUCMSCs transplantation. In vitro, antioxidants pretreatment of the hUCMSCs can decrease cell apoptosis through the MAPK-PKC-Nrf2 pathway. In vivo, antioxidants pretreatment can accelerate the host hepatic regenerative process and improve the expansion efficiency of stem cells. In this research, the antioxidant we used is edaravone which is now widely used in clinic. Thus, our study has answered how the oxidative stress affects stem cell transplantation efficiency and puts forward a clinical antioxidant which can improve the endogenous antioxidant level of stem cells.

Human umbilical cord-derived mesenchymal stromal cells ameliorated motor defects of 6-OHDA-induced rat model of Parkinson's disease

// Fabin Han 1, 6, * , Chao Chen 1, * , Wei Wang 1, * , Hao Song 1 , Sen Li 1 , Jing Duan 1 , Xianjie Lu 1 , Shichao Wu 1 , Nan Zhang 1 , Qingfa Chen 1 , Yan Wang 1 , Shuwei Liu 2 , Chongluo Fu 3 , Chengbiao Lu 4 and Paul Lu 5 1 Centre for Stem Cells and Regenerative Medicine, The Institute for Tissue Engineering &amp; Regenerative Medicine, Liaocheng University/The Liaocheng People&rsquo;s Hospital, Liaocheng, Shandong, China 2 Department of Human Anatomy, Shandong University, Jinan, Shandong, China 3 College of Biological Science, Liaocheng University, Liaocheng, Shandong, China 4 Department of Neurobiology, Xinxiang Medical College, Xinxiang, Henan, China 5 Department of Neurosciences, University of California at San Diego, La Jolla, CA, USA 6 The Institute for Translational Medicine, The Second Affiliated Hospital, Shandong University, Jinan, Shandong, China * These authors have contributed equally to this work Correspondence to: Fabin Han, email: fhan2013@126.com Keywords: Parkinson's disease; dopamine neuronal differentiation; umbilical cord-derived mesenchymal stromal cell; transplantation; gerotarget Received: July 29, 2017&emsp;&emsp;&emsp;&emsp; Accepted: January 01, 2018&emsp;&emsp;&emsp;&emsp; Published: January 02, 2018 ABSTRACT Cell therapy have a great potential for the treatment of neurodegenerative disease, such as Parkinson's disease (PD). Human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) have been reported to have multipotent differentiation ability. However, the therapeutic impact and mechanisms of dopamine neuronal differentiation from hUC-MSCs in PD are not determined. Here we developed a new protocol to induce dopamine neuron conversion from hUC-MSCs by addition of the growth cocktail containing noggin, CHIR99021, SHH, FGF8, TGF&beta;, GDNF, and BDNF. Then we transplanted the hUC-MSCs and the growth factor cocktail into the lesion side of the midbrain of 6-OHDA lesioned rat model of PD. The effects of hUC-MSC transplantation on the dopaminergic neuronal differentiation and motor behaviors of the rats were investigated. We found that in the presence of these molecules, the cultured hUC-MSCs showed a high efficient DA neuronal conversion in vitro . In combination with the growth cocktail, grafted hUC-MSCs also showed a highly efficient DA neuronal conversion in the midbrain of 6-OHDA lesioned rats. Both the grafted hUC-MSCs and the differentiated TH-positive neurons survived in 6-OHDA lesioned rats during the post-grafting period of 16 weeks. The hUC-MSCs-derived TH-positive neurons displayed the same electrophysiological profile as DA neurons in vivo . More importantly, rats with transplanted hUC-MSCs showed progressive improvements in motor behaviors compared to controls from weeks 4 to 16 post-grafting. These results demonstrated the efficacy and usefulness of the growth cocktail in combination with hUC-MSC transplantation in 6-OHDA lesioned rats and provided a promising cell-based treatment strategy for the PD patients.

VEGF gene transfected umbilical cord mesenchymal stem cells transplantation improve the lower limb vascular lesions of diabetic rats

BackgroundPrevious studies had explored the therapeutic effect of hUC-MSCs transplantation on ischemia; in this study, we further assessed the effectiveness of VEGF over-expressed hUC-MSCs cells transplantation on vascular proliferation in lower limb ischemia model in type 2 diabetic rats. MethodshUC-MSCs cells were over-expressed with VEGF, and transplanted to lower limb ischemia rats model of type 2 diabetes. ResultsVEGF over-expression increased the hUC-MSCs cells proliferation activity and VEGF secretion. VEGF gene transfected hUC-MSCs transplantation increased VEGF expression at a high level throughout 4weeks in skeletal muscle tissues of rats. Importantly, the vascular proliferation and blood perfusion of VEGF over-expressed hUC-MSCs transplanted limb were significantly improved compared with those of control group. The expression levels of ERK, AKT, MMP2 and MMP9 in VEGF over-expressed hUC-MSCs transplantation group increased dramatically compared with control group, while TIMP1 and TIMP2 expression had no significant change. ConclusionVEGF over-expressed hUC-MSCs transplantation was more effective to stimulate angiogenesis and increase blood perfusion than the simply hUC-MSCs transplantation, as maybe a new choice to improve the lower limb vascular lesions of diabetics.

Effect of human umbilical cord mesenchymal stem cells transplantation on nerve fibers of a rat model of endometriosis.

Endometriosis is a common, benign, oestrogen-dependent, chronic gynaecological disorder associated with pelvic pain and infertility. Some researchers have identified nerve fibers in endometriotic lesions in women with endometriosis. Mesenchymal stem cells (MSCs) have attracted interest for their possible use for both cell and gene therapies because of their capacity for self-renewal and multipotentiality of differentiation. We investigated how human umbilical cord-MSCs (hUC-MSCs) could affect nerve fibers density in endometriosis. In this experimental study, hUC-MSCs were isolated from fresh human umbilical cord, characterized by flow cytometry, and then transplanted into surgically induced endometriosis in a rat model. Ectopic endometrial implants were collected four weeks later. The specimens were sectioned and stained immunohistochemically with antibodies against neurofilament (NF), nerve growth factor (NGF), NGF receptor p75 (NGFRp75), tyrosine kinase receptor-A (Trk-A), calcitonin gene-related peptide (CGRP) and substance P (SP) to compare the presence of different types of nerve fibers between the treatment group with the transplantation of hUC-MSCs and the control group without the transplantation of hUC-MSCs. There were significantly less nerve fibers stained with specific markers we used in the treatment group than in the control group (p<0.05). MSC from human umbilical cord reduced nerve fiber density in the treatment group with the transplantation of hUC-MSCs.

Mesenchymal stem cells from the human umbilical cord ameliorate fulminant hepatic failure and increase survival in mice

Cell therapy has been promising for various diseases. We investigated whether transplantation of human umbilical cord mesenchymal stem cells (hUCMSCs) has any therapeutic effects on D-galactosamine/lipopolysaccharide (GalN/LPS)-induced fulminant hepatic failure in mice. hUCMSCs isolated from human umbilical cord were cultured and transplanted via the tail vein into severe combined immune deficiency mice with GalN/LPS-induced fulminant hepatic failure. After transplantation, the localization and differentiation of hUCMSCs in the injured livers were investigated by immunohistochemical and genetic analyses. The recovery of the injured livers was evaluated histologically. The survival rate of experimental animals was analyzed by the Kaplan-Meier method and log-rank test. hUCMSCs expressed high levels of CD29, CD73, CD13, CD105 and CD90, but did not express CD31, CD79b, CD133, CD34, and CD45. Cultured hUCMSCs displayed adipogenic and osteogenic differentiation potential. Hematoxylin and eosin staining revealed that transplantation of hUCMSCs reduced hepatic necrosis and promoted liver regeneration. Transplantation of hUCMSCs prolonged the survival rate of mice with fulminant hepatic failure. Polymerase chain reaction for human alu sequences showed the presence of human cells in mouse livers. Positive staining for human albumin, human alpha-fetoprotein and human cytokeratin 18 suggested the formation of hUCMSCs-derived hepatocyte-like cells in vivo. hUCMSC was a potential candidate for stem cell based therapies. After transplantation, hUCMSCs partially repaired hepatic damage induced by GalN/LPS in mice. hUCMSCs engrafted into the injured liver and differentiated into hepatocyte-like cells.

Enhanced angiogenesis promoted by human umbilical mesenchymal stem cell transplantation in stroked mouse is Notch1 signaling associated

Cellular therapy has provided hope for restoring neurological function post stroke through promoting endogenous neurogenesis, angiogenesis and synaptogenesis. The current study was based on the observation that transplantation of human umbilical cord mesenchymal stem cells (hUCMSCs) promoted the neurological function improvement in stroked mice and meanwhile enhanced angiogenesis in the stroked hemisphere. Grafted hUCMSCs secreted human vascular endothelial growth factor A (VEGF-A). Notch1 signaling was activated after stroke and also in the grafted hUCMSCs. To address the potential mechanism that might mediate such pro-angiogenic effect, we established a hUCMSC-neuron co-culture system. Neurons were subjected to oxygen glucose deprivation (OGD) injury before co-culturing to mimic the in vivo cell transplantation. Consistent with the in vivo data, co-culture medium claimed from hUCMSC-OGD neuron co-culture system significantly promoted the capillary-like tube formation of brain-derived endothelial cells. Moreover, coincident with our in vivo data, Notch 1 signaling activation was detected in hUCMSCs after co-cultured with OGD neurons as demonstrated by the up-regulation of key Notch1 signaling components Notch1 and Notch1 intercellular domain (NICD). In addition, OGD-neuron co-culture also increased the VEGF-A production by hUCMSCs. To verify whether Notch1 activation was involved in the pro-angiogenic effect, γ-secretase inhibitor N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT) was added into the co-culture medium before co-culture. It turned out that DAPT significantly prevented the Notch1 activation in hUCMSCs after co-culture with OGD neurons. More importantly, the pro-angiogenic effect of hUCMSCs was remarkably abolished by DAPT addition as demonstrated by inhibited capillary-like tube formation and less VEGF-A production. Regarding how Notch1 signaling was linked with VEGF-A secretion, we provided some clue that Notch1 effector Hes1 mRNA expression was significantly up-regulated by OGD-neuron co-culturing and down-regulated after additional treatment of DAPT. In summary, our data provided evidence that the VEGF-A secretion from hUCMSCs after being triggered by OGD neurons is Notch1 signaling associated. This might be a possible mechanism that contributes to the angiogenic effect of hUCMSC transplantation in stroked brain.

Human umbilical cord mesenchymal stem cells promote peripheral nerve repair via paracrine mechanisms.

Human umbilical cord-derived mesenchymal stem cells (hUCMSCs) represent a promising young-state stem cell source for cell-based therapy. hUCMSC transplantation into the transected sciatic nerve promotes axonal regeneration and functional recovery. To further clarify the paracrine effects of hUCMSCs on nerve regeneration, we performed human cytokine antibody array analysis, which revealed that hUCMSCs express 14 important neurotrophic factors. Enzyme-linked immunosorbent assay and immunohistochemistry showed that brain-derived neurotrophic factor, glial-derived neurotrophic factor, hepatocyte growth factor, neurotrophin-3, basic fibroblast growth factor, type I collagen, fibronectin and laminin were highly expressed. Treatment with hUCMSC-conditioned medium enhanced Schwann cell viability and proliferation, increased nerve growth factor and brain-derived neurotrophic factor expression in Schwann cells, and enhanced neurite growth from dorsal root ganglion explants. These findings suggest that paracrine action may be a key mechanism underlying the effects of hUCMSCs in peripheral nerve repair.

Human Umbilical Cord-Derived Mesenchymal Stem Cells Improve Learning and Memory Function in Hypoxic-Ischemic Brain-Damaged Rats via an IL-8-Mediated Secretion Mechanism Rather than Differentiation Pattern Induction

Background: MSCs are a promising therapeutic resource. Paracrine effects and the induction of differentiation patterns are thought to represent the two primary mechanisms underlying the therapeutic effects of mesenchymal stem cell (MSC) transplantation in vivo. However, it is unclear which mechanism is involved in the therapeutic effects of human umbilical cord-derived MSC (hUC-MSC) transplantation. Methods and Results: Based on flow cytometry analysis, hUC-MSCs exhibited the morphological characteristics and surface markers of MSCs. Following directed neural induction, these cells displayed a neuron-like morphology and expressed high levels of neural markers. All types of hUC-MSCs, including differentiated and redifferentiated cells, promoted learning and memory function recovery in hypoxic-ischemic brain damaged (HIBD) rats. The hUC-MSCs secreted IL-8, which enhanced angiogenesis in the hippocampus via the JNK pathway. However, the differentiated and redifferentiated cells did not exert significantly greater therapeutic effects than the undifferentiated hUC-MSCs. Conclusion: hUC-MSCs display the biological properties and neural differentiation potential of MSCs and provide therapeutic advantages by secreting IL-8, which participates in angiogenesis in the rat HIBD model. These data suggest that hUC-MSC transplantation improves the recovery of neuronal function via an IL-8-mediated secretion mechanism, whereas differentiation pattern induction was limited.

Human Umbilical Cord Mesenchymal Stem Cell Transplantation for the Treatment of Chronic Discogenic Low Back Pain

Background: Chronic low back pain is one of the major causes of disability and thus has a major socioeconomic impact. Intervertebral disc degeneration is the main cause of chronic low back pain. Treatment for chronic discogenic low back pain has traditionally been limited to either conservative management or surgical fusion. If conservative treatment fails, then surgical fusion is commonly considered. Current treatments are limited to treat the symptoms and not the underlying biologic alterations of the disc. Objective: Human umbilical cord tissue-derived mesenchymal stem cells (HUC-MSCs) contain stem cells and possess the ability to regenerate degenerative discs. Based on the results of previous in vitro and animal experiments, we conducted a preliminary study to test the feasibility and safety and to obtain an early indication for the therapeutic value of HUC-MSC transplantation in patients with chronic discogenic low back pain. Study Design: This is the first study involving treatment of chronic low back pain using HUCMSC transplantation. Setting: The study was performed at a spine center in China. Methods: Two patients with chronic discogenic low back pain were treated with HUC-MSC transplantation. An 11-point visual analog scale (VAS, 0 – 10) and Oswestry Disability Index (ODI, 0 – 100) were used to assess the back pain symptoms and the lumbar function, respectively. Results: After transplantation, the pain and function improved immediately in the 2 patients. The VAS and ODI scores decreased obviously during a 2-year follow-up period. Limitations: The shortcoming of this study is that it is a preliminary study with only 2 patients. Conclusion: The clinical outcomes indicated that HUC-MSC transplantation is a favorable alternative method for the treatment of chronic discogenic low back pain. Key words: Intervertebral disc degeneration, discogenic low back pain, chronic low back pain, lumbar discography, mesenchymal stem cells, human umbilical cord mesenchymal stem cells, transplantation

HUC-MSCs Transplantation Improve Microcirculatory Disturbance of Biliary Ischemia in Rabbit Model Evaluated By Contrast-Enchanced Ultrasonography (CEUS).

Objective To investigate efficacy of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) transplantation for improving microcirculatory disturbance of biliary ischemia in rabbit model. For the purpose of early and dynamic observation, contrast-enhanced ultrasonography (CEUS) was used. Methods 18 rabbits were divided randomly into 3 groups: the sham-operation group (SO group), ischemic-type biliary lesions (ITBLs) group, and hUC-MSCs transplantation + ITBLs group (MSCs+ITBL group). The model of ITBLs was established by clamping the hepatic artery and common bile duct for 2 hours. hUC-MSCs were transplanted though ear margin veins in 2h before operation in MSCs+ITBL group. the condition and microcirculatory of common and left lobe bile duct (CBD-LBD) was observed by CEUS at day 7 after operation. With the accompanying portal vein (PV) as reference, the following indexes were recorded: 1) the ratio of the caliber of CBD-LBD and the caliber of PV (PV/CBD-LBD); 2) the echotexture of the wall of CBD-LBD; 3) the thickness of the wall of CBD-LBD; 4) the integrity of the wall of CBD-LBD; and 5) the clarity of the CBD-LBD. Results The PV/CBD-LBD of the SO group was 3.47+0.23, and all the CBD-LBD (100%) showed iso-echoic, non-thickening and intact wall and clear lumen. Compared with SO group, that of ITBL group was 1.25+0.42(P<0.01). while in MSCs+ITBLs group, the PV/CBD-LBD of the SO group was 3.16+0.15 (compared with SO group and ITBLs group, P<0.01), and 5 rabbits showed iso-echoic, non-thickening and intact wall and clear lumen in CBD-LBD only 1 rabbits showed interrupt wall. All the CBD-LBD walls of the SO group showed linear hyper-enhancement during the arterial phase and hypo-enhancement in the portal venous and late phases. While in ITBL group, 4 rabbits's CBD-LBD walls showed hypo-enhancement in the arterial phase, and the other 2 persisted non-enhancement during the whole CEUS examination. In MSCs+ITBLs group, the CBD-LBD walls show the same performace in 5 rabbits as in SO group, only 1 rabbit showed hypoenhancement in the arterial phase. Conclusions hUC-MSCs transplantation can improve microcirculatory disturbance of biliary ischemia in rabbit model. As a new imaging modality, CEUS can clearly depict perfusion of the hilar bile duct.

Improvement of human umbilical cord mesenchymal stem cell transplantation on glial cell and behavioral function in a neonatal model of periventricular white matter damage

Periventricular white matter damage (PWMD) also termed periventricular leucomalacia in the preterm infant is of particular importance because no targeted therapy is presently available. Human umbilical cord mesenchymal stem cells (hUC-MSCs) have been studied in a variety of adult brain injury-related neurological disorders. Our aim was to determine whether hUC-MSC transplantation improves glial cell function in cerebral white matter and long-term behavioral function in a PWMD rat model. Rats on postnatal day (P) 3 underwent a permanent ligation of the left common carotid artery followed by 6% O2 for 4h. Immediately after the hypoxic–ischemic (HI), rats received a single intraperitoneal injection of hUC-MSCs, which were co-cultured with 5-bromodeoxyuridine (BrdU). BrdU+ cells in the brain were tested 24h after transplantation. Second, rats received hUC-MSC treatment once a day for 3 consecutive days. Glial cells (oligodendrocytes, astrocytes and microglia) were examined on 7 and 18 days post-HI, and behavioral outcomes were tested 27 days post-HI. Significantly, hUC-MSCs migrated mainly into the injured hemisphere. In addition, hUC-MSC treatment improved the long-term functional outcomes of rats, increased mature oligodendrocyte counts, and decreased the number of reactive astrocytes and activated microglia quantities after HI-induced damage in the premature brain. These results suggest that hUC-MSCs can pass through the blood–brain barrier and migrate towards the lesion site to improve brain damage. Therefore, hUC-MSCs have the potential to be utilized as a novel therapeutic strategy for PWMD.

Human Umbilical Cord Mesenchymal Stem Cells Transplantation Promotes Cutaneous Wound Healing of Severe Burned Rats

BackgroundSevere burns are a common and highly lethal trauma. The key step for severe burn therapy is to promote the wound healing as early as possible, and reports indicate that mesenchymal stem cell (MSC) therapy contributes to facilitate wound healing. In this study, we investigated effect of human umbilical cord MSCs (hUC-MSCs) could on wound healing in a rat model of severe burn and its potential mechanism.MethodsAdult male Wistar rats were randomly divided into sham, burn, and burn transplanted hUC-MSCs. GFP labeled hUC-MSCs or PBS was intravenous injected into respective groups. The rate of wound closure was evaluated by Image Pro Plus. GFP-labeled hUC-MSCs were tracked by in vivo bioluminescence imaging (BLI), and human-specific DNA expression in wounds was detected by PCR. Inflammatory cells, neutrophils, macrophages, capillaries and collagen types I/III in wounds were evaluated by histochemical staining. Wound blood flow was evaluated by laser Doppler blood flow meter. The levels of proinflammatory and anti-inflammatory factors, VEGF, collagen types I/III in wounds were analyzed using an ELISA.ResultsWe found that wound healing was significantly accelerated in the hUC-MSC therapy group. The hUC-MSCs migrated into wound and remarkably decreased the quantity of infiltrated inflammatory cells and levels of IL-1, IL-6, TNF-α and increased levels of IL-10 and TSG-6 in wounds. Additionally, the neovascularization and levels of VEGF in wounds in the hUC-MSC therapy group were markedly higher than those in other control groups. The ratio of collagen types I and III in the hUC-MSC therapy group were markedly higher than that in the burn group at indicated time after transplantation.ConclusionThe study suggests that hUC-MSCs transplantation can effectively improve wound healing in severe burned rat model. Moreover, these data might provide the theoretical foundation for the further clinical application of hUC-MSC in burn areas.

The potential of human umbilical cord-derived mesenchymal stem cells as a novel cellular therapy for multiple sclerosis.

Multiple sclerosis (MS) is a complex disease of neurological disability, affecting more than 300 out of every 1 million people in the world. The purpose of the study was to evaluate the therapeutic effects of human umbilical cord-derived mesenchymal stem cell (hUC-MSC) transplantation in MS patients. Twenty-three patients were enrolled in this study, and 13 of them were given hUC-MSC therapy at the same time as anti-inflammatory treatment, whereas the control patients received the anti-inflammatory treatment only. Treatment schedule included 1,000 mg/kg of methylprednisolone intravenously (IV) daily for 3 days and then 500 mg/kg for 2 days, followed by oral prednisone 1 mg/kg/day for 10 days. The dosage of prednisone was then reduced by 5 mg every 2 weeks until reaching a 5-mg/day maintenance dosage. Intravenous infusion of hUC-MSCs was applied three times in a 6-week period for each patient. The overall symptoms of the hUC-MSC-treated patients improved compared to patients in the control group. Both the EDSS scores and relapse occurrence were significantly lower than those of the control patients. Inflammatory cytokines were assessed, and the data demonstrated a shift from Th1 to Th2 immunity in hUC-MSC-treated patients. Our data demonstrated a high potential for hUC-MSC treatment of MS. This manuscript is published as part of the International Association of Neurorestoratology (IANR) special issue of Cell Transplantation.