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Background@#Preterm birth is an important health concern in countries with limited resources and healthcare access. Topical therapy may be effective for improving outcomes in preterm neonates whose skin barriers are compromised due to immaturity.@*Objectives@#To systematically review the topical VCO's effects in preterm infants on infection, mortality, and dermal maturity.@*Methodology@#Systematic review and meta-analysis of RCTs of topical VCO in preterm infants were conducted. Databases included PubMed, Google Scholar, Clinical trials.gov, Trip, Cochrane Library, and HERDIN. The risk of bias was assessed by two authors independently. RR with 95% CI was used for the pooled estimate of dichotomous outcomes including infection prevention, mortality reduction, and skin irritation. Mean differences with 95% CI were used for the pooled estimate of weight loss and NSCS.@*Results@#Of 110 records identified, 3 RCTs with 2440 patients were included. Prevention of infection had a trend toward VCO (RR = 0.90, [95% CI: 0.64, 1.27] while the results for mortality reduction were inconclusive (RR = 0.45, [95% CI: 0.06, 3.38]. NSC scores showed a beneficial trend toward VCO (RR = -0.03, [95% CI: -0.16, 0.09]. Both secondary outcomes of skin irritation and weight loss had inconclusive results.@*Conclusions@#This review showed the lack of evidence of the effectiveness of topical VCO in improving various outcomes in premature infants. The effects on infection prevention and dermal maturation were favorable. However, its effects on preventing mortality, skin irritation, and weight loss were inconclusive.
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Meta-Analysis , Infections , MortalityABSTRACT
OBJECTIVES: To investigate the molecular mechanism of edaravone (EDA) in improving the post-traumatic brain injury (TBI) dysfunction in learning and memory. METHODS: In vitro and in vivo TBI models were established using hydrogen peroxide (H2O2) treatment for hippocampal nerve stem cells (NSCs) and surgery for rats, followed by EDA treatment. WST 1 measurement, methylthiazol tetrazolium assay, and flow cytometry were performed to determine the activity, proliferation, and apoptosis of NSCs, and malondialdehyde (MDA), lactic dehydrogenase (LDH), and reactive oxygen species (ROS) detection kits were used to analyze the oxides in NSCs. RESULTS: Following EDA pretreatment, NSCs presented with promising resistance to H2O2-induced oxidative stress, whereas NSCs manifested significant increases in activity and proliferation and a decrease in apoptosis. Meanwhile, for NSCs, EDA pretreatment reduced the levels of MDA, LDH, and ROS, with a significant upregulation of Nrf2/antioxidant response element (ARE) signaling pathway, whereas for EDA-treated TBI rats, a significant reduction was observed in the trauma area and injury to the hippocampus, with improvement in memory and learning performance and upregulation of Nrf2/ARE signaling pathway. CONCLUSIONS: EDA, by regulating the activity of Nrf2/ARE signal pathway, can improve the TBI-induced injury to NSCs and learning and memory dysfunction in rats.
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Animals , Rats , Antioxidant Response Elements , Brain Injuries, Traumatic/physiopathology , Brain Injuries, Traumatic/metabolism , Brain Injuries, Traumatic/drug therapy , Edaravone/pharmacology , Learning/drug effects , Signal Transduction/drug effects , Reactive Oxygen Species/metabolism , Apoptosis/drug effects , Oxidative Stress/drug effects , NF-E2-Related Factor 2/metabolism , Memory/drug effectsABSTRACT
Objective:To investigate the effect of Buyang Huanwutang （BHT） on proliferation and differentiation in neural stem cells （NSCs） after oxygen-glucose deprivation/reoxygenation （OGD/R） injury. Method:NSCs isolated from the hippocampus of SD rats were cultured and randomly divided into a normoxia group， a model group， a BHT group， a rapamycin （Rapa） group， and a combination group ［autophagy inhibitor 3-methyladenine （3-MA） combined with BHT］. The 20% blank serum was used in the normoxia group， and 20% BHT-medicated serum in the BHT group. The doses of Rapa and 3-MA were 1 μmol·L<sup>-1</sup> and 5 mmol·L<sup>-1</sup>， respectively. The cells were subjected to OGD/R except those in the normoxia group. The cell morphology was observed under a light microscope. NSCs were confirmed by immunofluorescence detection of nestin expression. The viability and proliferation of NSCs were assessed by cell counting kit-8 （CCK-8） assay and 5-ethynyl-2-deoxyuridine （EdU） labeling， respectively. Furthermore， Ad-mCherry-GFP-LC3B fluorescence assay was performed to investigate autophagy. The effect of BHT on autophagy-related protein expression was detected by western blot assay. Brain derived neurotrophic factor （BDNF）， <italic>β</italic>-tubulin Ⅲ， and glial fibrillary acidic protein （GFAP） were evaluated by immunofluorescence assay. Result:OGD/R significantly reduced the cell viability of rat NSCs as compared with the normoxia group. Compared with the model group， the BHT group exhibited significantly improved viability of rat NSCs （<italic>P</italic><0.01）. BHT induced the production of autophagosomes in NSCs after OGD. The BHT group showed increased expression of microtuble-associated protein 1 light chain 3Ⅱ (LC3Ⅱ） and Beclin-1 （<italic>P</italic><0.05，<italic>P</italic><0.01） and slightly changed p62 compared with the normoxia group， and significantly up-regulated LC3Ⅱ and Beclin-1 （<italic>P</italic><0.05，<italic>P</italic><0.01） and down-regulated expression of p62 （<italic>P</italic><0.01） compared with the model group. The Rapa group had similar effect as the BHT group （<italic>P</italic><0.05，<italic>P</italic><0.01）， while the combination group inhibited the activity of autophagy （<italic>P</italic><0.01）. As indicated by the results of ad-mCherry-GFP-LC3B， compared with the normoxia group， the model group showed increased fluorescence intensity （<italic>P</italic><0.01）， and the BHT and Rapa groups could further increased the fluorescence intensity of autophagy （<italic>P</italic><0.01）， while the combination group inhibited autophagy activity （<italic>P</italic><0.01）. Immunofluorescence results revealed that compared with the normoxia group， the model group displayed significantly reduced positive cells of EdU， <italic>β</italic>-tubulin Ⅲ， GFAP， and BDNF （<italic>P</italic><0.01）， and the BHT and Rapa groups exerted similar protective and promoting effects （<italic>P</italic><0.05，<italic>P</italic><0.01）， while the combination group partially blocked the neuroprotection and differentiation ability of BHT （<italic>P</italic><0.05）. Conclusion:BHT pretreatment can effectively protect rat NSCs against OGD-induced injury and promoted proliferation and differentiation by up-regulating autophagy.
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Objective To investigate the effects of Bushen Huoxue acupuncture method on the behavioral changes and the proliferation of endogenous neural stem cells (NSCs) in hippocampus of Alzheimer disease (AD) model mice. Methods 18 male SAMP8 mice, seven months old, were randomly divided into acupuncture group, non-acupoint control group, and model control group. And another age-matched 6 male SAMR1 mice were prepared as normal control group. Mice in acupuncture group were intervened by acupuncture method in the acupoints of "Shenshu", "Baihui", "Xuehai", and "Geshu". Mice in non-acupoint control group were treated by stimulating the fixed non-point under the bilateral rib, while mice in model control group and normal control group were raised without special treatment but administered the stimulation of catching with the same time and the same stimulus intensity. All treatrment lasted for 8 weeks. After the intervention, the learning and memory abilitities and brain hippocampus Brd U positive cells of all groups were detected. Results Compared with the nomal control group, mice in the model control group had longer escape latency and less time spent in former platform quadrant (P＜0.05); the number of Brd U positive cells decreased significantly (P＜0.05). Compared with the model control group and the non-acupoint control group, mice in the acupuncture group had shorter escape latency and more time spent in former platform quadrant (P＜0.05); the number of Brd U positive cells increased significantly (P＜0.05). Conclusion Bushen Huoxue acupuncture method can improve the learning and memory abilities of SAMP8 mice AD model by inducing the proliferation of hippocampal endogenous NSCs.
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Objective To investigate the effect of berberine on proliferation of neural stem cells(NSCs)induced by hydrogen peroxide(H2O2). Methods NSCs from Sprague-Dawley rats were isolated and purified by suspension culture. Cells were divided into a control group,H2O2group(NSCs exposed to H2O2injury),berberine group(NSCs were incubated with berberine concentrations ranging from 0.5 to 20 μmol/L and exposed to H2O2), and DAPT(a blocker of the Notch signaling pathway)group. Cell viability was evaluated using the Cell Counting Kit-8 assay. Proliferation of NSCs was evaluated by a neurosphere formation assay and Ki67 protein expression. Expression of key proteins in the Notch signaling pathway(including notch1 and hes1)in response to berberine treatment or DAPT(a Notch inhibitor)was determined by Western blotting. Results Cell viability of NSCs was significantly increased by berberine compared with the H2O2group. The neurosphere growth assay showed that 5 or 10 μmol/L berberine increased NSC proliferation. The ratio of Ki67 +/DAPI cells and notch1 and hes1 protein expression increased significantly compared with the H2O2group. Conclusions Berberine treatment upregulates Notch signaling in NSCs,whereas DAPT attenuates these effects. Berberine is a drug that promotes NSC proliferation and exerts a protective effect on NSCs via the Notch signaling pathway.
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The paper uses the methods of bibliometrics and mathematical statistics to carry out statistical analysis on the scientific pa pers in Neural Stem Cells (NSCs) in 2007-2016 recorded by the SCI database from the aspects of distribution of years,countries,institutions,journals,funds and high-frequency keywords,and generally evaluates the development hotspots and application prospect of NSCs in the recent 10 years.
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Objective To study the relationship of proliferation of neural stem cells (NSCs) in SVZ and the expressions of c-jun and c-myc in rats with middle cerebral artery occlusion/reperfusion (MCAO/R) injury model administrated byDanlong XingnaoFormula.Methods The focal cerebral ischemia reperfusion injury models were prepared by longa method. Totally 150 male SD rats were randomly divided into sham-operation group, cerebral model group,Danlong XingnaoFormula low-, medium-, and high-dose groups. The treatment groups were given corresponding dose ofDanlong XingnaoFormula, while the sham-operation group and model group were given the same amount of distilled water 24 h after modeling by gavage, once a day, 7 days in a row. 1 d, 3 d and 7 d after reperfusion, modified Neurological Severity Scores (m-NSS) was used to grade neurologic impairment. 7 d after reperfusion taken to the SVZ brain tissue of ischemia side, Brdu immunohistochemical method was used to record the BrdU positive cells number. The hippocampal c-jun, c-myc mRNA and protein expressions were determined respectively by RT-qPCR method and Western blot method.Results Grades of neurologic impairment in others groups were improved obviously than sham-operation group (P<0.01); 3 d, and 7 d after reperfusion, grades of neurologic impairment inDanlong XingnaoFormula groups were obviously lower compared with model group (P<0.05,P<0.01). Brdu positive cell rates in others groups increased obviously compared with sham-operation group; Compared with model group, Brdu positive cell rates inDanlong XingnaoFormula groups increased obviously (P<0.01). The expressions of c-jun and c-myc protein and mRNA inDanlong XingnaoFormula groups improved obviously than sham-operation group and model group (P<0.01).ConclusionDanlong Xingnao Formula can improve the neural function after cerebral ischemia and stimulate the proliferation of NSCs, and the mechanism may be related to activating the expression of c-jun and c-myc and extending the duration.
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PURPOSE: Neural stem cells (NSCs) effectively reverse some severe central nervous system (CNS) disorders, due to their ability to differentiate into neurons. Agmatine, a biogenic amine, has cellular protective effects and contributes to cellular proliferation and differentiation in the CNS. Recent studies have elucidated the function of microRNA let-7a (let-7a) as a regulator of cell differentiation with roles in regulating genes associated with CNS neurogenesis. MATERIALS AND METHODS: This study aimed to investigate whether agmatine modulates the expression of crucial regulators of NSC differentiation including DCX, TLX, c-Myc, and ERK by controlling let-7a expression. RESULTS: Our data suggest that high levels of let-7a promoted the expression of TLX and c-Myc, as well as repressed DCX and ERK expression. In addition, agmatine attenuated expression of TLX and increased expression of ERK by negatively regulating let-7a. CONCLUSION: Our study therefore enhances the present understanding of the therapeutic potential of NSCs in CNS disorders.
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Agmatine , Biogenic Amines , Cell Differentiation , Cell Proliferation , Central Nervous System , MicroRNAs , Neural Stem Cells , Neurogenesis , NeuronsABSTRACT
PURPOSE: Neural stem cells (NSCs) effectively reverse some severe central nervous system (CNS) disorders, due to their ability to differentiate into neurons. Agmatine, a biogenic amine, has cellular protective effects and contributes to cellular proliferation and differentiation in the CNS. Recent studies have elucidated the function of microRNA let-7a (let-7a) as a regulator of cell differentiation with roles in regulating genes associated with CNS neurogenesis. MATERIALS AND METHODS: This study aimed to investigate whether agmatine modulates the expression of crucial regulators of NSC differentiation including DCX, TLX, c-Myc, and ERK by controlling let-7a expression. RESULTS: Our data suggest that high levels of let-7a promoted the expression of TLX and c-Myc, as well as repressed DCX and ERK expression. In addition, agmatine attenuated expression of TLX and increased expression of ERK by negatively regulating let-7a. CONCLUSION: Our study therefore enhances the present understanding of the therapeutic potential of NSCs in CNS disorders.
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Agmatine , Biogenic Amines , Cell Differentiation , Cell Proliferation , Central Nervous System , MicroRNAs , Neural Stem Cells , Neurogenesis , NeuronsABSTRACT
Aim Toclarifytheeffectofacteosideon proliferation of neural stem cells (NSCs ) from adult mice,as well as the involved signaling pathway.Meth-ods NSCswereisolatedfromthesubventricularzone (SVZ)of adult C57BL/6 mice,then identified by im-munofluorescence staining with Nestin,the marker of NSCs.NSCs were exposed to acteoside (5,10,20,40μmol·L-1 )in absence of mitogen(EGF/bFGF)for 24 h.We employed CCK8 assay to detect NSCs viability and BrdU staining to identify NSCs proliferation.We performed Western blot to quantify the expression level ofp-AktinducedbyacteosideonNSCs.Results With-out mitogen,acteoside increased NSCs proliferation by activating p-Akt,which can be blocked by LY294002, the inhibitor of PI3K/AKT signaling pathway.Conclu-sion ActeosidepromotestheproliferationofNSCsfrom adult mice by activating PI3K/AKT pathway.
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The purpose of the study was to investigate the impact of rat cytomegalovirus(RCMV) infection on the development of the nervous system in rat embryos,and to evaluate the involvement of Wnt signaling pathway key molecules and the downstream gene neurogenin 1(Ngn1) In RCMV infected neural stem cells(NSCs).Infection and control groups were established,each containing 20 pregnant Wistar rats.Rats in the infection group were inoculated with RCMV by intraperitoneal injection on the first day of pregnancy.Rat E20 embryos were taken to evaluate the teratogenic rate.NSCs were isolated from E13 embryos,and maintained in vitro.We found:1) Poor fetal development was found in the infection group with low survival and high malformation rates.2) The proliferation and differentiation of NSCs were affected.In the infection group,NSCs proliferated more slowly and had a lower neurosphere formation rate than the control.The differentiation ratio from NSCs to neurons and glial cells was significantly different from that of the control,showed by immunofluorescence staining.3) Ngn1 mRNA expression and the nuclear β-catenin protein level were significantly lower than the control on day 2 when NSCs differentiated.4) The Morris water maze test was performed on 4-week pups,and the infected rats were found worse in learning and memory ability.In a summary,RCMV infection caused abnormalities in the rat embryonic nervous system,significantly inhibited NSC proliferation and differentiation,and inhibited the expression of key molecules in the Wnt/β-catenin signaling pathway so as to affect NSCs differentiation.This may be an important mechanism by which RCMV causes embryonic nervous system abnormalities.
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Objective To study the effects of embryonic neural stem cells transplantion on trauma of red nucleus neu-rons of the rats with spinal cord injury.Methods NSCs in logarithmic phage were labeled with BrdU,a Sprague Dawley rat mode of spinal cord injury (SCI) was developed with electrocircuit control spinal cord injuring device.Thirty SD rats were randomly divided into three groups: sham group,SCI group and NSC group.The NSCs were trans-planted into injured site three days after SCI.Then NSCs labeled with Brdu were detected by immunohistochemisty,rubrospinal tract (RST) neurons were labeled by retrograde transport of the horseradish peroxidase (HRP) from the lesion site,which were taken by damaged axons and remained in the neurons,then the labeled red nucleus (RN) neurons were counted.Hind limb function of experimental rats was evaluated by a blinder observer using BBB open field locomotion rating score.Results BrdU positive NSCs were detected in the spinal cord after transplantation,the number of RST neurons labeled by HRP in NSC group was more than that in SCI group (P <0.01),the BBB score of NSC group was higher than SCI group (P <0.01).Conclusion The transplanted NSCs can survive in the injured site of spinal cord and protect RN,then promote more remarkably functional recovery after SCI.
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Objective To look for a small molecular neurotrophin which could promote production and/or differentiation of neural stem cells (NSCs). Methods 1.Embryo hippocampal NSCs were cultured in vitro . 2. The neurospheres were identified by antibodies of bromodeoxyuridine (BrdU), glial fibrillary acid protein (GFAP), microtubule associated protein 2 (MAP2) and Galactocerebroside (GalC). 3. The cells were divided into four groups, which were control group, FBS treated group, trans-sequence of APP 5-mer peptide treated group, APP 5-mer peptide treated group. The morphology of NSCs was observed in above four groups. 4. Cell counts, detection of clone information rate and diameter of clone were done to study the effect of APP 5-mer peptide on production of NSCs. 5. In addition, we also detected the MTT metablism rates in all groups. Results 1. The NSCs formed neurospheres and grew in floating. They were BrdU-positive. GFAP-positive, MAP2-positive and GalC-positive cells appeared after FBS were added into the medium. 2. The morphology of NSCs was not changed in APP 5-mer peptide treated group and trans-sequence group compared with the control group. 3. The cell number increased in APP 5-mer peptide treated group as compared with the control group. There were no apparent differences between the control group and the trans-sequence treated group. 4. The clone formation rate and diameters of neurospheres increased in APP 5-mer peptide treated group. 5. The MTT metabolism rate increased in APP 5-mer peptide treated group. Conclusion APP 5-mer peptide could promote the production of embryo hippocamal NSCs in vitro. APP 5-mer peptide doesn't promote the differentiation of embryo hippocamal NSCs in vitro.
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In order to examine the effect of IGF-1 on neurogenesis after focal cerebral ischemia in young and olderrats.IGF-1 was applied by transventricular injection one day before operation of permanent middle cerebral artery occlusion(MCAO).Incontrast,the control groups were treated with vehicle. Methods: Bromodeoxyuridin (BrdU) was used as a marker of the proliferating cells,and PSA-NCAM as a marker of neural precursor cells,BrdU-labeled cells immunoreacted With MAP2 as a marker of differentiated neural precursor cells 28d after MCAO.Rats were administered With BrdU 6 days after MCAO.Immunohistochemistry was used to de-tect the expression of BrdU and PSA-NCAM immtmoreactivity in cortex and hippocampus 7 days and 28 days after MCAO.double im-munohistochemistry was used to detect the expression of MAP2 of BrdU.labeled cells 28d after MCAO.Results:The number of BrdU-labeled cells and PSA-NCAM positive cells increased 5.1 fold and 3.2 7d after 1GF-1 infusion in older rats.and 5.5 fold and 3.2 fold in young r ts.compared with vehicle.treated group(p>0.05).The residual rate of BrdU.labeled cells were 79.2%and 75.1% respectively inyoungandolderrats (p>0.05),incontraryt 077.1% and 52.3%(p<0.05)invehicle.treatedgroups 28d after MCAO.BrdU/MAP2 positive cells increased after IGF.1 infusion with more clear effect in young group(p<0.05),compared with vehicle.treated group.Conclu-sion:Our results indicated that IGF-1 pretreatment induced the neurogenesis,ameliorated the survival of post-proliferation cells in MCAO rats and induced neural precursor cells to differentiate into neuron.Our finding will be helpful to developing therapeutic applica-t-ion of IGF.1 after brain injury in older patients.
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In order to examine the effect of IGF-1 on neurogenesis after focal cerebral ischemia in young and olderrats.IGF-1 was applied by transventricular injection one day before operation of permanent middle cerebral artery occlusion(MCAO).Incontrast,the control groups were treated with vehicle. Methods: Bromodeoxyuridin (BrdU) was used as a marker of the proliferating cells,and PSA-NCAM as a marker of neural precursor cells,BrdU-labeled cells immunoreacted With MAP2 as a marker of differentiated neural precursor cells 28d after MCAO.Rats were administered With BrdU 6 days after MCAO.Immunohistochemistry was used to de-tect the expression of BrdU and PSA-NCAM immtmoreactivity in cortex and hippocampus 7 days and 28 days after MCAO.double im-munohistochemistry was used to detect the expression of MAP2 of BrdU.labeled cells 28d after MCAO.Results:The number of BrdU-labeled cells and PSA-NCAM positive cells increased 5.1 fold and 3.2 7d after 1GF-1 infusion in older rats.and 5.5 fold and 3.2 fold in young r ts.compared with vehicle.treated group(p>0.05).The residual rate of BrdU.labeled cells were 79.2%and 75.1% respectively inyoungandolderrats (p>0.05),incontraryt 077.1% and 52.3%(p<0.05)invehicle.treatedgroups 28d after MCAO.BrdU/MAP2 positive cells increased after IGF.1 infusion with more clear effect in young group(p<0.05),compared with vehicle.treated group.Conclu-sion:Our results indicated that IGF-1 pretreatment induced the neurogenesis,ameliorated the survival of post-proliferation cells in MCAO rats and induced neural precursor cells to differentiate into neuron.Our finding will be helpful to developing therapeutic applica-t-ion of IGF.1 after brain injury in older patients.
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0.05).The transfected NSCs were confirmed to have the latent ability of multi-directional differentiation.Conclusion:NSCs can express HIF-1? and GFP steadily after transfected with the recombinant adenovirus and the bionomics is normal.
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@#ObjectiveTo investigate bone marrow mesenchymal stem cells(BMSC) express brain derived neurotrophic factor(BDNF) and nerve growth factor(NGF) and their protective effect for neural stem cells (NSCs).MethodsBMSC were obtained from rat tibiae and femurs and centrifuged with Ficoll. The passage 3 cells were chosen to make immunocytochemical stain for CD44, CD71 and CD45. The expression of BDNF and NGF was detected in BMSC with RT-PCR, as well as in the media with ELISA. The media that cultured BMSC were collected as BMSC condition media. NSCs were obtained from cerebral cortex of new-born rat and cultured in vitro. After different ratio of BMSC condition midia were added, NSCs were induced to apoptosis with heat-shock, then NSCs were dyed with Annexin V-FITC/PI apoptosis kit and apoptosis rates were tested with flow cytometry. ResultsBMSC were CD44(+), CD45(-), CD71(+) and expressed BDNF and NGF mRNA. BDNF and NGF could be tested in the media of cultured BMSC and increase with cultured time. BMSC condition media could reduce the ratio of heat-induced apoptosis of NSCs, and more BMSC condition media showed better effect. ConclusionBMSC can express neurotrophic factores and protect neural stem cells from heat-induced apoptosis.
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@#ObjectiveTo investigate the effects of fluid percussion injury(FPI) on survival and differentiation of transplanted human embryonic neural stem cells (HNSCs) in rats. MethodsThe HNSCs were separated from the cerebral cortex of the 8-week-old fetal and were cultured in DMEM/F12 combinated with EGF, bFGF and LIF. The rat models of FPI were made with fluid percussion system. The HNSCs labeled with BrdU were transplanted into the injured zone 24 hours after brain injury, then the rats were killed at the 1st and 4th week post-transplanted stages, and the brain slices were stained with immunocytochemistry. The GFAP, MAP-2, and BrdU positive cells were investigated.ResultsThe transplanted HNSCs migrated to the whole brain, and differentiated into GFAP and MAP-2 positive cells. MAP-2 positive cells were observed at 1 week post-transplanted stage, on the contrary, more GFAP positive cells were discovered 4 weeks after transplantation. Part of the HNSCs migrated to the choroids plexus of the lateral ventricle and microvessels. ConclusionThe transplanted HNSCs survive in the injured zone, and differentiate into astrocytes gradually during the recovery. The host devours part of the HNSCs.
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@#ObjectiveTo investigate an effective method to isolate neural stem cells(NSCs).MethodsNSCs were dissociated by digestion with trypsin, EDTA and different doses of Dispase,and serum-free culture techniques and immunohistochemistry techniques were used to verifying the dissociated.ResultsA lot of single neural stem cells were obtained by using Dispase to digest neurosphere, and the cells could keep its structure and morphology.ConclusionIt is an ideal method by using Dispase to digest neurosphere for isolating NSCs.
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@#ObjectiveTo study the mechanism of differentiation of mesenchymal stem cells(MSCs) into neuron-like cells in vitro.MethodsMSCs of Wistar rats were separated and cultured, and then induced with DMSO and BHA in vitro. The specific marking proteins of neurons, glia and neural stem cells were detected before preinduction, at 24h after preinduction, at 6h, 24h, and 48h after neuronal induction.ResultsAfter the inducement, many MSCs turned into bipolar,multipolar and taper,and then intersected as network structure. Nestin was strong positive at 6h after neuronal induction, and decreased at 24h, 48h after the induction. NeuN was present at 6 h after neuronal induction, and increased at 24h, 48h after the induction.ConclusionMSCs can be induced into neural stem cells(NSCs) at first, and then differentiate into neuron-like cells in vitro.