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【Objective】 To investigate the effect of Polymerized human cord hemoglobin (PolyCHb) on the sensitivity of hepatocellular carcinoma grafts to lumvalatinib in nude mice. 【Methods】 Hep3B hepatoma cells were subcutaneously transplanted in 18 nude mice to establish tumor graft model. Mice were randomly divided into 3 groups: control group (the saline 90 mg·kg-1·d-1), monotherapy group (Lenvatinib10 mg·kg-1·d-1), and sensitized group (Lenvatinib mg·kg-1·d-1, polyCHB 600 mg/kg twice a week) for 28 days. The tumor volume was measured regularly and the growth curve was drawn. On day 29, the nude mice were sacrificed, the tumor was stripped and weighed, and the pathomorphological differences of each group were evaluated by HE section staining. The expression levels of hypoxia-inducing factor (HIF-1α), CD34, VEGF, CD44, MMP-9, and Glut-1 in tumor tissues of each group were determined by immunohistochemistry. The content of reactive oxygen species (ROS) in tumor tissues of each group was determined by dihydroethyl ingot method. 【Results】 The tumor growth rate and tumor volume in the sensitized group decreased significantly compared with the control group and the solo drug group. On day 29, the tumor volumes of the control group, the monotherapy group and the sensitization group were (2 076.46±350.25)mm3, (1 035.96±84.16)mm3 and (892.66±104.46)mm3, respectively. Tumor weight was (1.61±0.52)g, (0.45±0.10)g, and (0.34±0.13)g, respectively. Immunohistochemical score of HIF-1α was 75±23 vs 45±18 vs 18±11, VEGF was 52±8 vs 67±16 vs 35±4, CD34 was 40±7 vs 50±13 vs 28±7, CD44 was 37±15 vs 30±7 vs 15±3, Glut-1 was 74±41 vs 51±30 vs 14±18, MMP-9 was 51±7 vs 62±20 vs 33±3, respectively(P<0.05). The malignant degree of the sensitized group was decreased by HE section staining, which was significantly lower than that of the solo drug group and the control. The ROS content in the sensitized group was higher than that in the solo drug group and the control. 【Conclusion】 PolyCHb can reduce the expression of HIF-1α and its downstream pathway related molecules by increasing oxygenation of hepatocellular carcinoma tissues in nude mice, delay tumor growth and reduce tumor volume in a certain period, thus increase the therapeutic effect of lenvalatinib on hepatocellular carcinoma grafts in tumor bearing nude mice models.
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BACKGROUND: A large number of studies mainly concern the proliferation effect of mesenchymal stem cells on hematopoietic stem cells in vitro and that bone marrow mesenchymal stem cell transplantation can reduce the death of hematopoietic cells caused by irradiation, increase the survival of bone marrow cells and repair hematopoiesis, while few of them investigate the repair of human umbilical cord blood mesenchymal stem cells transplantation on bone marrow hematopoiesis injury. OBJECTIVE: To explore the repair of hematopoietic microenvironment of bone marrow by human umbilical cord blood mesenchymal stem cells. METHODS: Male BALB/c mice were randomly divided into three groups. The mice in experimental group and control group were irradiated with total dose of 6-Gy X-ray to establish a mouse model of bone marrow hematopoietic injury. The normal group contained untreated normal mice. In the experimental group, CM-DiL labeled human umbilical cord blood mesenchymal stem cells were injected into the tail vein of each mouse at 5×106 (0.2 mL). The control group and the normal group received normal saline 0.2 mL through the tail vein. The peripheral blood hematology and bone marrow hematopoietic microenvironment repair were observed at 1, 5, 7, 14 and 21 days after cell transplantation. RESULTS AND CONCLUSION: Peripheral blood condition: At 1, 5 and 7 days after transplantation, the leucocyte, platelet, erythrocyte count and hemoglobin concentration in the experimental group and control group decreased progressively compared with the normal group. The most obvious decrease occurred on day 7. The trilineage recovered on day 14 after transplantation, basically returned to normal on day 21 after transplantation. Compared with the experimental group, the decrease of the trilineage in the control group was more obvious. The recovery was obvious faster in the experimental group than in the control group on day 14 after transplantation. Bone marrow smears: Bone marrow smears showed that the hematopoietic function was inhibited in the experimental group and the control group at 1, 5, 7, and 14 days after transplantation, especially on day 7. Bone marrow proliferation recovered on day 14 after transplantation. It was better in the experimental group than in the control group. On day 21 after transplantation, the hematopoietic function of bone marrow of mice in the experimental group and the control group recovered, and there was no difference between the experimental group and the control group compared with the normal group. Bone marrow pathological section: Bone marrow pathological sections showed that at 1, 5, 7, and 14 days after transplantation, the hematopoietic function of bone marrow in the experimental group and the control group was inhibited. On day 14 after transplantation, the bone marrow hematopoietic function of the experimental group and the control group began to recover, but the bone marrow proliferation of the experimental group was better than that of the control group. On day 21 after transplantation, there was no difference in the bone marrow proliferation between the experimental and the control groups and the normal group. The results suggested that human umbilical cord blood mesenchymal stem cells can promote the recovery of hematopoietic function of bone marrow.
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Wharton's jelly mesenchymal stem cells (WJ-MSCs) are a class of stem cells with high differentiative potential, an immuno-privileged status and easy access for collection, which raise no legal or ethical issues. WJ-MSCs exhibit several features of embryonic stem cells, both in the phenotypic and genetic aspects, with only a few differences, such as a shorter doubling time and a more extensive ex vivo expansion capacity. WJ-MSCs have immunomodulatory properties, involving both innate and adaptive immune responses. This review focuses on the role of WJ-MSCs in the management of graft-versus-host disease (GvHD), a life-threatening complication of the allogenic transplantation of hematopoietic stem cells. Different studies documented the beneficial effect of the infusion of WJ-MSCs, even when not fully HLA identical, in patients with severe GvHD, refractory to standard treatment. Finally, we summarized current ongoing clinical trials with WJ-MSCs and their potential in regenerative medicine.
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Humans , Embryonic Stem Cells , Ethics , Graft vs Host Disease , Hematopoietic Stem Cells , Immunomodulation , Mesenchymal Stem Cells , Regenerative Medicine , Stem Cells , Umbilical Cord , Wharton JellyABSTRACT
Objective@#To explore the effects of shear stress on morphology, adhesion and proliferation of human umbilical cord blood mesenchymal stem cells(hUC-MSCs).@*Methods@#The hUC-MSCs were cultured in vitro until confluence and then placed in a flow system.The cells were subjected to different shear stress (1, 2, 3, 4 dye/cm2) for 2 hours and 6 hours, and no shear stress treatment cells served as a static control.The morphological changes of hUC-MSCs in different groups were analyzed by phase contrast microscopy and immunofluorescence.The mRNA expression levels of intercellular adhesion molecule-1 (ICAM-1) and Ki67 were detected by real-time PCR.@*Results@#Compared with the static control group, the hUC-MSCs cells were arranged in the direction of fluid after treated with shear stress.The immunofluorescence results showed that the cytoskeletal protein F-actin filaments was prolonged after shear stress.The cytoskeleton was further elongated in the 2 dye/cm2 for 6 hours group when compared with 2 dye/cm2 for 2 hours group, and the cytoskeleton was loosened when time extended to 6 hours.Real-time PCR results showed that the relative expressions of ICAM-1 mRNA and Ki67 mRNA in the static control group and different gradient shear stress groups were significantly different, with significant differences among them (F=17.141, P=0.000; F=11.336, P=0.001). The relative expression of ICAM-1 mRNA in the 1, 2, 3, 4 dye/cm2 shear stress group was 2.74±0.32, 9.77±1.19, 6.70±0.92 and 5.69±0.72, respectively, which was significantly higher than 1.00±0.28 in the static control group, with significant differences between them (all at P<0.05). The relative expression of Ki67 mRNA in the 3 dye/cm2 and 4 dye/cm2 shear stress group was 0.39±0.09 and 0.04±0.02, respectively, which was significantly lower than 1.00±0.24 in the static control group, with statistically significant differences between them (both at P<0.05).@*Conclusions@#After treated with fluid shear stress, hUC-MSCs are arranged in the direction of fluid.Shear stress can promote the adhesion of hUC-MSCs.As the increase of shear stress intensity, cell proliferation is inhibited.
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OBJECTIVE@#To investigate the effect of Piwil2-induced cancer stem-like cell (Piwil2-iCSC)-derived exosomes on the proliferation,migration and invasion of human umbilical cord blood-derived mesenchymal stem cells (hucMSCs).@*METHODS@#Piwil2-iCSC-derived exosomes were isolated by ultracentrifugation and identified using transmission electron microscopy,nanoparticle tracking analysis and Western blotting.Exosome uptake assay was used to identify the pathway that Piwil2-iCSCderived exosomes utilized.HucMSCs were divided into control group,PBS intervention group and exosome intervention group,and CCK-8 assay,wound healing assay,Transwell assay,Western blotting and cell karyotype analysis were used to observe the proliferation,migration,invasion,expression levels of MMP2 and MMP9 proteins,and chromosome structure of hucMSCs.@*RESULTS@#The diameter of Piwil2-iCSC-derived exosomes ranged from 50 nm to 100 nm,and most of them were oval or spherical capsules rich in CD9,CD63 and Piwil2 proteins.Exosomal uptake assay showed that the exosomes executed theirs functions after entering the cells.Compared with the control cells and PBS-treated cells,hucMSCs treated with the exosomes showed significantly increased number of proliferating cells (<0.05) with accelerated healing rate (<0.05 at 24 h;<0.01 at 48 h),increased invasive cells (<0.01),enhanced protein expressions of MMP2(<0.05 PBS group;<0.01 control group) and MMP9(<0.05),but their karyotype still remained 46XY without any abnormalities.@*CONCLUSIONS@#Piwil2-iCSC-derived exosomes can promote the proliferation,migration and invasion but does not cause cancer-like heterogeneity changes in hucMSCs.
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Humans , Argonaute Proteins , Cell Movement , Physiology , Cell Proliferation , Physiology , Exosomes , Physiology , Fetal Blood , Cell Biology , Karyotyping , Mesenchymal Stem Cells , Pathology , Neoplasm Invasiveness , Neoplastic Stem Cells , Umbilical Cord , Wound HealingABSTRACT
BACKGROUND AND OBJECTIVES: Sensorineural hearing loss (SNHL) in children is associated with neurocognitive morbidity. The cause of SNHL is a loss of hair cells in the organ of Corti. There are currently no reparative treatments for SNHL. Numerous studies suggest that cord blood mononuclear cells (human umbilical cord blood, hUCB) allow at least partial restoration of SNHL by enabling repair of a damaged organ of Corti. Our objective is to determine if hUCB is a safe treatment for moderate to severe acquired SNHL in children. SUBJECTS AND METHODS: Eleven children aged 6 months to 6 years with moderate to severe acquired SNHL were treated with intravenous autologous hUCB. The cell dose ranged from 8 to 30 million cells/kg body weight. Safety was assessed by measuring systemic hemodynamics during hUCB infusion. Infusion-related toxicity was evaluated by measuring neurologic, hepatic, renal and pulmonary function before and after infusion. Auditory function, auditory verbal language assessments and MRI with diffusion tensor imaging (DTI) were obtained before and after treatment. RESULTS: All patients survived, and there were no adverse events. No infusionrelated changes in hemodynamics occurred. No infusion-related toxicity was recorded. Five subjects experienced a reduction in auditory brainstem response (ABR) thresholds. Four of those 5 subjects also experienced an improvement in cochlear nerve latencies. Comparison of MRI with DTI sequences obtained before and after treatment revealed increased fractional anisotropy in the primary auditory cortex in three of five subjects with reduced ABR thresholds. Statistically significant (p < 0.05) reductions in ABR thresholds were identified. CONCLUSIONS: TIntravenous hUCB is feasible and safe in children with SNHL.
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Child , Humans , Anisotropy , Auditory Cortex , Body Weight , Cochlear Nerve , Diffusion Tensor Imaging , Evoked Potentials, Auditory, Brain Stem , Fetal Blood , Hair , Hearing Loss, Sensorineural , Hemodynamics , Magnetic Resonance Imaging , Mesenchymal Stem Cells , Organ of Corti , Umbilical CordABSTRACT
Objective To induce the differentiation of human umbilical cord blood MSCs into osteoblasts in vitro,and to study the method of inducing MSCs to differentiate into osteoblasts under specific microenvironment.Methods MSCs was obtained from human umbilical vein,and isolated by density gradient method.The morphological changes of MSCs were observed by using dexamethasone,beta sodium phosphate and vitamin C.The proliferation and differentiation of MSC in cord blood were studied by means of optical microscope,transmission electron microscope and alkaline phosphatase staining.The expression of bone morphogenetic protein 2 mRNA in human umbilical cord blood MSCs after osteogenic was inducted by RT-PCR.Results After the umbilical cord blood MSCs were differentiated into osteoblastic cells in microenvironment,fusiform cells became polygonal,irregular shape,local cells presented overlapping growth.After 10 days,the cells gradually presented square,crystal particles of high refraction,and began to show the characteristics of osteoblasts.The expression of bone morphogenetic protein 2 mRNA was positive in alkaline phosphatase staining and alizarin red staining.Conclusion Human umbilical cord blood MSCs can be induced into osteoblasts in vitro,which is an ideal seed cell for bone tissue engineering.
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Recent advances have shown the direct reprogramming of mouse and human fibroblasts into induced neural stem cells (iNSCs) without passing through an intermediate pluripotent state. Thus, direct reprogramming strategy possibly provides a safe and homogeneous cellular platform. However, the applications of iNSCs for regenerative medicine are limited by the restricted availability of cell sources. Human umbilical cord blood (hUCB) cells hold great potential in that immunotyped hUCB units can be immediately obtained from public banks. Moreover, hUCB samples do not require invasive procedures during collection or an extensive culture period prior to reprogramming. We recently reported that somatic cells can be directly converted into iNSCs with high efficiency and a short turnaround time. Here, we describe the detailed method for the generation of iNSCs derived from hUCB (hUCB iNSCs) using the lineage-specific transcription factors SOX2 and HMGA2. The protocol for deriving iNSC-like colonies takes 1~2 weeks and establishment of homogenous hUCB iNSCs takes additional 2 weeks. Established hUCB iNSCs are clonally expandable and multipotent producing neurons and glia. Our study provides an accessible method for generating hUCB iNSCs, contributing development of in vitro neuropathological model systems.
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Animals , Humans , Mice , Fetal Blood , Fibroblasts , In Vitro Techniques , Methods , Neural Stem Cells , Neuroglia , Neurons , Regenerative Medicine , Transcription Factors , Umbilical CordABSTRACT
Stem cell therapies are administered during the acute phase of stroke to preserve the penumbral tissues from ischemic injury. However, the effect of repeated cell therapy during the acute phase remains unclear. In this study, we investigated and compared the functional outcome of single (two days post-injury) and repeated (two and nine days post-injury) treatment with human umbilical cord derived mesenchymal stem cells (hUCB-MSCs) after middle cerebral artery occlusion (MCAO). The rotarod and limb placement tests were utilized to investigate functional outcomes, while infarct volume and tissue damage were measured by immunofluorescent staining for neovascularization, neurogenesis, apoptosis, and inflammation in the penumbral zones. We observed notable motor dysfunction and a significant decrease in infarcted brain volume, as well as increases in neurons and vessels in both single and repeated hUCB-MSC treatments compared to the control group. Interestingly, repeated administration of hUCB-MSCs was not found to elicit additional or synergistic improvements over monotherapy. This study suggests that a clearer understanding of the therapeutic window after stroke will facilitate the development of more efficient treatment protocols in the clinical application of stem cell therapy.
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Animals , Humans , Rats , Apoptosis , Brain , Brain Ischemia , Cell- and Tissue-Based Therapy , Clinical Protocols , Extremities , Infarction, Middle Cerebral Artery , Inflammation , Ischemia , Mesenchymal Stem Cells , Neurogenesis , Neurons , Stem Cells , Stroke , Umbilical CordABSTRACT
Objective:To observe the changes of blood glucose,insulin and dipeptidyl peptidase-Ⅳ(DPP-Ⅳ/CD26)on type 2 diabetes mellitus in rabbits after HUCBSC( human umbilical cord blood stem cells) transplantation. Methods:18 rabbits were randomly divided into normal control group (6 rats,Group C) and diabetic model group (12 rats). After preparation model of type 2 diabetes,and 6 rats of them were treated with HUCBSC ( CD45+,CD34-) transplantation by ear vein transfusion ( Group A) ,and 6 rats were treated with PBS(Group B). All three groups of rabbits were fed for 4 weeks,and the blood glucose was monitored every day,and the level of blood insulin and DPP-IV/CD26 were measured every week. Results:The negative expression rate of CD34 in HUCBSC was 96. 5%. The positive expression rate of CD45 in HUCBSC was 100%. Compared with non transplantation group,the blood glucose and DPP-IV/CD26 in the umbilical cord blood stem cell transplantation group were gradually decreased,and insulin level was gradually increased, the difference was statistically significant (P<0. 01). Conclusion:HUCBSC were round or oval,with adherent growth,HUCBSC trans-plantation can significantly reduce blood glucose, increase insulin secretion, reduce the level of DPP-IV/CD26, the immunological phenotype of HUCBSC was CD45+,CD34-,thus providing a new theoretical basis for the clinical treatment of diabetes and its complica-tions.
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The expression of immunogenic markers after differentiation of umbilical cord blood (UCB)-derived mesenchymal stem cells (MSC) has been poorly investigated and requires extensive in vitro and in vivo testing for clinical application. The expression of human leukocyte antigen (HLA) classes on UCB-derived MSC was tested by Fluorescence-activated cell sorting analysis and immunocytochemical staining. The undifferentiated MSC were moderately positive for HLA-ABC, but almost completely negative for HLA-DR. The MSC differentiated to chondrocytes expressed neither HLA-ABC nor HLA-DR. The proliferation of MSC was not significantly affected by the allogeneic lymphocytes stimulated with concanavalin A. The responder lymphocytes showed no significant decrease in proliferation in the presence of the MSC, but the apoptosis rate of the lymphocytes was increased in the presence of MSC. Taken together, these findings indicate that UCB-derived MSC differentiated to chondrocytes expressed less HLA class I and no class II antigens. The MSC showed an immunomodulatory effect on the proliferation and apoptosis of allogeneic lymphocytes. These data suggest that the differentiated and undifferentiated allogeneic MSC derived from umbilical cord blood can be a useful candidate for allogeneic cell therapy and transplantation without a major risk of rejection.
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Humans , Apoptosis , Cell- and Tissue-Based Therapy , Chondrocytes , Concanavalin A , Fetal Blood , Flow Cytometry , Histocompatibility Antigens Class II , HLA-DR Antigens , In Vitro Techniques , Leukocytes , Lymphocytes , Mesenchymal Stem Cells , Umbilical CordABSTRACT
Objective To investigate the clinical efficacy of umbilical blood stem cell transplantation (UCBSCT) on the treatment of hepatitis B liver cirrhosis. Methods Forty-eight patients with hepatitis B liver cirrhosis were enrolled and divided into the treatment group and the control group. There were 25 patients in the treatment group , who received UCBSCT treatment based on conventional liver protection treatment and 23 patients in the control group , who received conventional liver protection treatment. The changes of liver function , coagulation function, clinical symptoms, signs and side effects were studied before the treatment and at 2, 4, 12 and 24 weeks post-treatment. Results The levels of albumin, cholinesterase, and prothrombin activity in the treatment group were higher than those before treatment and were higher than those in the control group. The parameters in the control group were not significantly changed before and after the treatment (P > 0.05). The levels of alanine aminotransferase,aspartate aminotransferase,total bilirubin in both two groups were not significantly changed before and after the treatment (P > 0.05). After 4-week treatment,the differences on improvement of appetite , lacking in strength , abdominal distension , ascites were statistically significant in the treatment group compared with the control group (P < 0.05). No adverse reactions were observed in all groups. Conclusion UCBSCT on the treatment of hepatitis B liver cirrhosis is safe and reliable.
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Objective To investigate the effect of transplantation of human umbilical cord blood CD34+cells on spinal cord injury. Methods CD34+cells were separated from fresh human umbilical cord blood by magnetic cell sorting. Ninety-six female Wistar rats were injured at T10 by IMPACTOR MODEL-Ⅱ, and then randomly assigned to three groups:Cyclo?sporin A (CsA)+Dexamethasone (Dex) treated group (Ⅰ, n=32), local transplantation of cells+CsA+Dex treated group (Ⅱ) at the first day after operation (DAO 1, n=32), local transplantation of cells+CsA+Dex treated group (Ⅲ) at DAO 6 (n=32). BBB locomotor scoring system was used to assess the recovery of the lower limbs. The survival and neural differentiation of transplanted cells at the injury site were observed by double immunofluorescence. The tissue vitality at the injury site was ob?served by 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) staining, and the blood vessel density was observed by infusing mixture of Chinese ink and glutin followed by HE staining. Results BBB score at DAO 8-56 was significantly higher inⅡgroup than that of other two groups (P<0.05). TTC staining showed that the proportion of decreased vitality area was signifi?cantly smaller inⅡgroup than that of other two groups (P<0.01). The result of gelatin ink perfusion showed that the blood vessel density at the injury site was significantly bigger inⅡgroup than that of other two groups (P<0.01). There were more survival transplanted cells inⅡgroup than those of III group (per visual field, 7.51 ± 1.00 vs 5.51 ± 0.89,t=6.051, P<0.01). All the transplanted cells didn’t differentiate into neural cells. Conclusion Human umbilical cord blood CD34+cells can promote the recovery of the lower limbs after spinal cord injury by repairing blood vessels to increase tissue vitality at the in?jury site in rats.
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Objective:To explore the effect of intratracheal transplantation human umbilical cord blood mesenchymal stem cells on pulmonary interstitial fibrosis by bleomycin in mice,compare the treatment in pulmonary fibrosis of intratracheal transplantation with tail vain injection human umbilical cord blood mesenchymal stem cells.Methods:The second generation of HUCBMSCs were cultured to the fourth generation.Sixty specific pathogen free male Kunming mice were randomly divided into 4 groups:negative control group ( Cont group) ,bleomycin group( BLM group) ,HUCBMSCs transplantation groupⅠ( MⅠgroup) and HUCBMSCs transplantation groupⅡ(MⅡ group),each group 15 mice.Pulmonary fibrosis models were induced by bleomycin via intratracheal perfusion in the latter three groups.Twenty four hours after model establishment,5-bromo-2-deoxyuridine( Brdu) marked HUCBMSCs were poured in trachea in MⅠgroup ,the same were injected into tail vein in MⅡ group.At the 7th,14th,28th day,5 mice in each group were executed re-spectively.The morphological changes of the lung tissues were observed by HE staining and Masson staining.The localization and distribution of human umbilical cord blood mesenchymal stem cells were determined by the method of immunohistochemistry.The hydroxyproline contents were measured by alkali hydrolysis assay.The protein levels of transforming growth factor β-1 ( TGF-β1 ) and smooth muscle alpha-actin(α-SMA) were detected by Western blot.Results:In the two mesenchymal stem cell transplantation groups, there were Brdu marked cells at the 7th,14th,28th days in lung tissue.The alveolitis and fibrosis in lung of the two mesenchymal stem cell transplantation groups were milder than which of the the bleomycin group(PMⅠ,MⅡgroup>Cont group(P0.05).Conclusion:The colonization of human umbilical cord blood mesenchymal stem cells can be seen in the damaged tissue via intratracheal transplantation which can alleviate pulmonary fibrosis in mice caused by bleomycin.
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Objective:To explore the effects of compound Danshen dripping pills (CDDP) and CDDP combined with transplantation of human umbilical cord blood cells (HUMNCs) on the inlfammatory response, oxidative stress, myocardial cell apoptosis and cardiac function, and also to investigate the possible mechanisms of the combined therapy in the acute myocardial infarction (AMI). Methods:Rabbit model of AMI successfully established by ligation of the letf anterior coronary artery (LAD). Forty rabbits were randomly divided into 4 groups (n=10 per group):a control group, injected with 0.5 mL of saline in 24 h atfer AMI and then gavaged with 5 mL of saline daily;a CDDP group, injected with saline 0.5 mL atfer AMI and then gavaged with CDDP (270 mg/d) daily;a transplantation group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs [labeled with green fluorescent protein (GFP)] and then gavaged with 5 mL of saline daily;a combined group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs (labeled with GFP) and then gavaged with CDDP (270 mg/d) daily. Cardiac function index such as left ventricular fractional shorting (LVFS) and ejection fraction(LVEF) were measured by echocardiography;the pathological changes were observed by HE staining and the white blood cells in the myocardium were determined by light microscopy. hTe superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in myocardium were detected by nitrotetrazolium blue chloride (NBT) and thiobarbituric acid colorimetric measurement respectively. hTe number of transplanted cells in the myocardium was examined by GFP positive cells counted with lfuorescence microscopy. Results:1) Compared with the control group (at 1 or 4 week), LVEF and LVFS were signiifcant improved in the CDDP group, the transplantation group and the combined groups (all P Conclusion:The intravenous transplantation of HUMNCs combined with the CDDP in the treatment of rabbits with AMI could increase the survival rate of transplanted cells and inhibit the myocardial cell apoptosis, therefore improve the heart function. hTe possible mechanism of the combined treatment may be involved in the inhibition of the inlfammatory response and oxidative stress in the myocardium following AMI.
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10.3969/j.issn.2095-4344.2013.23.002
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Cell transplantation is a promising experimental treatment for spinal cord injury. The aim of the present study was to evaluate the efficacy of mononuclear cells from human umbilical cord blood in promoting functional recovery when transplanted after a contusion spinal cord injury. Female Wistar rats (12 weeks old) were submitted to spinal injury with a MASCIS impactor and divided into 4 groups: control, surgical control, spinal cord injury, and one cell-treated lesion group. Mononuclear cells from umbilical cord blood of human male neonates were transplanted in two experiments: a) 1 h after surgery, into the injury site at a concentration of 5 x 10(6) cells diluted in 10 µL 0.9 percent NaCl (N = 8-10 per group); b) into the cisterna magna, 9 days after lesion at a concentration of 5 x 10(6) cells diluted in 150 µL 0.9 percent NaCl (N = 12-14 per group). The transplanted animals were immunosuppressed with cyclosporin-A (10 mg/kg per day). The BBB scale was used to evaluate motor behavior and the injury site was analyzed with immunofluorescent markers to label human transplanted cells, oligodendrocytes, neurons, and astrocytes. Spinal cord injury rats had 25 percent loss of cord tissue and cell treatment did not affect lesion extension. Transplanted cells survived in the injured area for 6 weeks after the procedure and both transplanted groups showed better motor recovery than the untreated ones (P < 0.05). The transplantation of mononuclear cells from human umbilical cord blood promoted functional recovery with no evidence of cell differentiation.
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Animals , Female , Humans , Rats , Fetal Blood/cytology , Leukocytes, Mononuclear/transplantation , Spinal Cord Injuries/surgery , Cell Differentiation , Nerve Regeneration , Rats, Wistar , Recovery of Function , Transplantation, HeterologousABSTRACT
Objective To investigate the effect of human umbilical cord blood stem cells on flash visual evoked potentials (F-VEP) of the traumatic optic neuropathy rats.Methods Forty-eight Sprague-Dawley rats were randomly divided into an injury group (Group A) and 3 treatment groups (Groups B,C,and D).A traumatic optic neuropathy model was built in Group A,and the rats in Groups B,C,and D were injected with the neurotrophic factor,human umbilical cord blood stem cells,and the mixture of the neurotrophic factor and human umbilical cord blood stem cells,respectively.F-VEP was recorded in both eyes of rats at the 1st h,1st week,2nd week,3rd week,and 4th week after the optic nerve injury.Results At all time points,there were significant difference in the wave latency and amplitude between Group A and normal control eyes (P＜0.01).The differences of the wave latency and amplitude between Group A and Groups B,C,and D were statistically significant at various time points after the injury except for the wave latency at the 1st h post-operation (P＞0.05).The amplitude in Group D was higher while the latency was shorter than those of Group B at all time points since the 1st week (P＜0.05).The comparisons at the same point in the remaining treatment groups were not significantly different (P＞0.05).Conclusion The mixture of human umbilical cord blood stem cells and neurotrophic factor has a promotion effect for the recovery of F-VEP of optic nerve in traumatic optic neuropathy in rats to some degrees.
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BackgroundOptic nerve injury lead to apoptosis of retinal ganglion cells ( RGCs ), and its mechanism of apoptosis is endoplasmic reticulum stress (ERS). So, decreasing of ERS may protect the injury of RGCs. ObjectiveThe present study was to investigate the mechanisms of endoplasmic reticulum stress(ERS) and the protective effects of human umbilical cord blood stem cells on partial optic nerve crush injury. MethodsThe optical nerves were crushed with a 40 g clip by holding for 60 seconds to establish the partial optical nerve injury model in the left eyes of 102 SPF SD rats,and 10 μl of mRNA and 10 μl of nerve growth factor were injected into the vitreous immediately after the establishment of the model. The morphological changes of retinal ganglion cells(RGCs) were examined under the light microscope after 3,7,14,21 and 28 days and the RGCs number was calculated. The apoptosis rates of RGCs were detected by the TUNEL technique after 3, 12,24,45,72 hours and 1 week. The expression levels of GRP78 mRNA and CHOP mRNA were detected by reverse transcription polymerase chain reation (RT-PCR). This procedure followed the Regulations for the Administration of Affair Concerning Experimental Animals by Hunan Provincial Science and Technology Committee.Results The number of RGCs was significantly decreased with the prolongation of time of optical nerve injury in the model injury group,whereas the number of RGCs in the human cord blood cells group was reduced at a slower rate( Ftime =20. 100,P =0. 007 ). At various time points after the injection of human cord blood cells, the survival of RGCs was evidently increased in comparison with the model group(P＜0. 01 ). The apoptosis rate of RGCs was considerably elevated with injury time prolongation both in the model group and human cord blood cells group,but no apoptosis was seen from 3-24 hours after operation,and only a small amount of apoptotic cells were found in the human cord blood cells group from 48 hours through 1 week than in the model group(P＜0. 01 ). In the human cord blood cells group,GRP78 mRNA level was significantly higher and the CHOP mRNA level was significantly lower than those in the injury group at identical time points(P＜0. 01 ). ConclusionsIn the rat optic nerve partial crush model,ERS induces the apoptosis of RGCs. Human umbilical cord blood stem cells can protect RGCs from ERS injury by inhibiting apoptosis.
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Therapy with bone marrow-derived cells has been used in ischemic patients with reported success. The aim of this study was to determine the therapeutic efficacy of fresh and frozen human umbilical cord blood cells (hUCB) in Wistar rats submitted to permanent occlusion of the left coronary artery. Three hours after myocardial infarction, 2 x 10(7) hUCB cells or vehicle were administered by intramyocardial injection. The animals were divided into five groups: control (N = 10), sham operated (N = 10), infarcted that received vehicle (N = 9), infarcted treated with cryopreserved hUCB (N = 7), and infarcted treated with fresh hUCB (N = 5). Cardiac function was evaluated by electrocardiogram (ECG) and echocardiogram (ECHO) before cell therapy, and by ECG, ECHO, cardiopulmonary test, and left ventricular pressure measurements 3 weeks later. After 3 weeks, both groups treated with hUCB still had Q wave present in L1, âQRS >90° and reduced shortening fraction (less than 50 percent). In addition, cardiac indexes of left ventricular contractility and relaxation were 5484 ± 875 and -4032 ± 643 mmHg (cryopreserved hUCB) and 4585 ± 955 and -2862 ± 590 mmHg (fresh hUCB), respectively. These values were not statistically different from those of saline-treated animals. Cardiopulmonary exercise test profile was typical of infarcted hearts; exercise time was about 14 min and maximal VO2 was 24.77 ± 5.00 mL·kg-1·min-1. These data show that hUCB therapy did not improve the cardiac function of infarcted animals or prevent cardiac remodeling.