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【Objective】 To investigate the effect and mechanism of achyranthes bidentata polysaccharides （ABPS） on the differentiation of bone mesenchymal stem cells （BMSCs） into nucleus pulposus-like cells. 【Methods】 Five 4-week-old SD rats were sacrificed and their BMSCs were isolated and purified by repeated adherent method. The third-generation BMSCs were treated with different concentrations of ABPS for 48 h. The cell viability was detected by MTT method. The highest concentration of ABPS was selected for subsequent experiments. According to different treatment methods， cells were divided into control group （conventional culture）， ABPS group （200 mg/L of ABPS）， Notch1 group （transfected with Notch1）， Notch1 negative control group （transfected with Notch1 negative control）， and ABPS+Notch1 group （200 mg/L of ABPS was added after transfection with Notch1）. After 14 days of induction， the cell survival rate was measured by MTT method. CollagenⅡ was detected by immunohistochemical staining. The mRNA and protein expression levels of collagenⅡ， aggrecan， Notch1 and Hes1 were detected by qPCR and Western Blotting respectively. The glucosaminoglycan （GAG） was detected by alcian blue method on the 1st， 4th， 7th， 10th and 14th day of cell culture. 【Results】 After BMSCs were treated with 400 mg/L of ABPS for 48 h， the cell survival rate was significantly lower than that in the control group （P<0.05）. When the concentration was ≤200 mg/L， ABPS had no significant toxic effect on the growth of BMSCs. After 14 days of induction， compared with the control group， A value， GAG content in the medium， the expressions of collagen II and aggrecan mRNA and protein in the ABPS group were increased， and the expressions of Notch1 and Hes1 mRNA and protein were decreased （P<0.05）. The relative cell viability， GAG content in the medium， the expressions of collagen II and aggrecan mRNA and protein in the Notch1 group were decreased， and the expressions of Notch1 and Hes1 mRNA and protein were increased （P<0.05）. ABPS+Notch1 could partially reverse the inhibitory effect of Notch1 overexpression on the differentiation of BMSCs into nucleus pulposus-like cells. 【Conclusion】 ABPS can promote the differentiation of BMSCs into nucleus pulposus-like cells， and the mechanism may be related to the inhibition of Notch1 pathway.
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Objective:To explores the improvement in survival mechanism of a rat model of enterocoelia heterotopic heart transplant with rat bone marrow mesenchymal stem cells( BMSCs) IDO-overexpressing. Methods:IDO-overexpressing rat BMSCs were produced through transfection of rat BMSCs with IDO gene carried by the lentiviral vector GV308. A rat model of enterocoelia heterotopic heart transplantation was established. This rat model received a cell treatment via its tail veins, as follows: ①Echocardiography was employed to detect the functional changes in the transplanted heart.②The fluorescence intensity of the different parts of the transplanted heart was evaluated using a body imaging system for small living animals.③Receptors rat spleens cells were obtained and used for a flow cytometric detection of the expression levels of CD40,CD86,CD80,MHCⅡ,CD274,CD45RA,CD45RA+CD45RB,and Treg cells. ④A transplanted heart was obtained after injection to evaluate inflammatory cell infiltration through HE staining.⑤Liquid phase chips were used to detect changes in the serum factors IL-1ɑ,IL-4,IL-1β,IL-2,IL-10,IFN-γ,IL-18,TGFβ1, TGFβ2 and TGFβ3 in after injection cells. Results:①After the rat heterotopic heart transplantation model and the corresponding cell treatment were established,after over-expressed IDO-BMSCs treatment 2 days the EF and FS were higher in the transplanted heart than other groups.②The fluorescence intensity of the parts of the transplanted heart was highest in the IDO-BMSC overexpression group as revealed by small animal living body evaluation. ③Two days after the interventions, spleen cells in the over-expressed IDO-BMSCs group showed reduced expression levels of CD40,CD86,CD80,MHCⅡ,CD45RA,CD45RA+CD45RB and increased expression levels of CD274 and Treg cells as revealed by flow cytometry.④Liquid phase chips were used to examine the serum obtained from each group 2 days after the intervention,and the results showed that the expression levels of IL-1α,IL-4,IL-1β,IL-2,IFN-γand IL-18 in the IDO-BMSC overexpression group decrease. By contrast,the expression levels of IL-10,TGFβ1,TGFβ2 and TGFβ3 increase. HE staining results demonstrate that inflammatory cell infiltration was lower in IDO-BMSC overexpression group than in other groups. Conclusion:IDO-overexpressing BMSCs improve the survival of a transplanted heart through effective adjustment of immune DC and T cells,as well as cell factors.
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Objective:To explores the improvement in survival mechanism of a rat model of enterocoelia heterotopic heart transplant with rat bone marrow mesenchymal stem cells( BMSCs) IDO-overexpressing. Methods:IDO-overexpressing rat BMSCs were produced through transfection of rat BMSCs with IDO gene carried by the lentiviral vector GV308. A rat model of enterocoelia heterotopic heart transplantation was established. This rat model received a cell treatment via its tail veins, as follows: ①Echocardiography was employed to detect the functional changes in the transplanted heart.②The fluorescence intensity of the different parts of the transplanted heart was evaluated using a body imaging system for small living animals.③Receptors rat spleens cells were obtained and used for a flow cytometric detection of the expression levels of CD40,CD86,CD80,MHCⅡ,CD274,CD45RA,CD45RA+CD45RB,and Treg cells. ④A transplanted heart was obtained after injection to evaluate inflammatory cell infiltration through HE staining.⑤Liquid phase chips were used to detect changes in the serum factors IL-1ɑ,IL-4,IL-1β,IL-2,IL-10,IFN-γ,IL-18,TGFβ1, TGFβ2 and TGFβ3 in after injection cells. Results:①After the rat heterotopic heart transplantation model and the corresponding cell treatment were established,after over-expressed IDO-BMSCs treatment 2 days the EF and FS were higher in the transplanted heart than other groups.②The fluorescence intensity of the parts of the transplanted heart was highest in the IDO-BMSC overexpression group as revealed by small animal living body evaluation. ③Two days after the interventions, spleen cells in the over-expressed IDO-BMSCs group showed reduced expression levels of CD40,CD86,CD80,MHCⅡ,CD45RA,CD45RA+CD45RB and increased expression levels of CD274 and Treg cells as revealed by flow cytometry.④Liquid phase chips were used to examine the serum obtained from each group 2 days after the intervention,and the results showed that the expression levels of IL-1α,IL-4,IL-1β,IL-2,IFN-γand IL-18 in the IDO-BMSC overexpression group decrease. By contrast,the expression levels of IL-10,TGFβ1,TGFβ2 and TGFβ3 increase. HE staining results demonstrate that inflammatory cell infiltration was lower in IDO-BMSC overexpression group than in other groups. Conclusion:IDO-overexpressing BMSCs improve the survival of a transplanted heart through effective adjustment of immune DC and T cells,as well as cell factors.
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Objective To investigate the effect of hyperoxia on the expressions of cysteinyl aspartate specific proteinase -3(Caspase -3)and proliferating cell nuclear antigen (PCNA)in bone marrow mesenchymal stem cells (BMSCs).Methods Primary BMSCs from SD rats were cultured in vitro,BMSCs grew to 70% -80% degrees of fu-sion and then were randomly divided into room air group and hyperoxia exposure group.Each group was divided into 5 sub -groups,and cultured for 0,3,6,1 2,and 24 h respectively.Morphology investigation with inverted phase contrast microscope was adopted.The expressions of Caspase -3 and PCNA protein levels were detected by Western blot. Results (1 )As time wenty by,compared with room air group,the gap between cells increased,some of the cells be-came circular,and cell detachment and floating appeared in the hyperoxia -exposure group (>1 2 h)compared with room air group.(2)As time went on,compared with the room air group,the expression levels of Caspase -3 protein in the hyperoxia -exposure group were increased,and the difference was significant after 6 hours of culture (0.27 ± 0.04 vs 0.1 4 ±0.02,t =5.03,P =0.007).(3)Compared with room air group,the PCNA levels of the hyperoxiaexpo-sure group(6 h)decreased,and the difference in PCNA protein expression levels was significant after 6 hours of culture (0.27 ±0.04 vs 0.38 ±0.04,t =3.37,P =0.028).Conclusions Hyperoxia exposure increases Caspase -3 expres-sion levels and decreases PCNA expression,which may affect the proliferation and apoptosis of BMSCs.
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[Abstract ] Objective To explore the effective method of induction of Schwann cell-like differentiation in bone mesenchymal stem cell (BMSC)of adult rat in vitro.Methods Primary culture of Schwann cell and isolated culture of BMSC were separately conducted.According to different induction methods,the cells were divided into chemical induction group and co-culture induction group.The growth of Schwann cell and BMSC was observed under light microscope.These two kinds of cells were identified by immunofluorescence staining [detecting Schwann cell marker proteins:glial fibrillary acidic protein (GFAP) antibody and S-100 antibody] and flow cytometry.The shape and growth of cells in two groups were dynamically observed by light microscope.The induced differentiation was evaluated with immunofluorescence staining at 3 rd day after co-culture induction in the co-culture induction group and at 4 h and 1 st day after chemical induction in the chemical induction group.Results In the chemical induction group,the BMSC appeared typical Schwann cell-like morphology.The positive expression of GFAP antibody appeared at 4 h after preliminary induction.Meanwhile,the positive expression rate of GFAP and S-100 antibody was (80.9 ± 3.5)% and (59.0 ±1.1 )% at 1 st day after induction.The induced BMSC began to die at 2nd day after chemical induction and most of the induced BMSC had died at 3 rd day after chemical induction.At 3 rd day after co-culture induction,few induced BMSC showed obvious morphological changes like those in chemical induction group.The positive expression rate of GFAP and S-100 antibody was (89.8 ±2.4)% and (80.9 ±1.7)%. The positive expression rate of GFAP and S-100 antibody in the co-culture induction group was higher than those in the chemical induction group and the difference had statistical significance (all in P <0.01).Conclusions The co-culture induction not only has obvious effect on Schwann cell-like differentiation in BMSC,but also promotes the survival and proliferation of BMSC.Thus,co-culture induction is more safe and effective than chemical induction.
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Objective To study the regulation mechanism of bone mesenchymal stem cell (MSC)combined co-translation of islets in differentiation of Follicular Helper T cell (Tfh),and its roll on immunotolerence induction in non-obese diabetic (NOD) mice transplantation model.Methods The NOD mice were divided into 4 groups:Group A with islet transplantation alone;Group B with MSC co-transplantation with islets (MSC:0.5 × 106);Group C with MSC co-transplantation with islets (MSC:2 × 106);Group D with MSC co-transplantation with islets (MSC:3 × 106).ELISA was used to test the expression level of diabetes autoantibody GAD65Ab and IAA.Tfh cell count was detected by FACS.Results The survival time of transplantation groups was much longer in MSCs co-transplantation group than islet-alone group;the level of GAD65Ab,IAA and Tfh cell count were much lower in MSCs co-transplantation group than islet-alone group.Conclusion MSC may protect the islet transplants by regulating the Tfh cell differentiation.
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Objective To explore the effect of acupoint injection of bone marrow mesenchymal stem cells (BMSCs)on hemodynamics of rat model with acute myocardial infarction(AMI). Methods Sixty Sprague-Dawley (SD)rats were randomly divided into sham group,model group,myocardium injection BMSCs group and acupoint injection BMSCs group(each n=15). The left anterior descending coronary artery(LAD)was ligated to establish a rat model of AMI. After the rat model was successfully established for 72 hours,0.2 ml BMSCs(1×1010/L)were transplanted by a micro-quantity syringe at 6 points in equal amount in the LAD blood-supply area and its periphery in the myocardial injection group,while in the acupoint injection group,0.3 ml BMSCs(1×1010/L)was transplanted at each of the following acupoints:Xinshu,Zhiyang and Tanzhong. Four weeks after AMI,polyethylene tubing was inserted into the right carotid artery to measure the hemodynamics,at the same time animals were sacrificed,and the heart was take out to calculate the heart mass index(HMI)and left ventricular mass index(LVMI). Results One, 3,5 and 4 rats were respectively dead in the sham group,model group,myocardium injection group,and acupoint injection group during the experimental period. Compared with the sham group,the left ventricular systolic pressure (LVSP,mm Hg,1 mm Hg=0.133 kPa),the maximum rate of increase of left ventricular pressure(+dp/dt max, mm Hg/s), the maximum rate of decrease of left ventricular pressure(-dp/dt max,mm Hg/s), the maximum logarithmic change rate of left ventricular pressure〔(dp/dt)?P-1max,s-1〕,HMI(mg/g),LVMI were significantly decreased,and left ventricular end-diastolic pressure(LVEDP,mm Hg),heart rate(HR,bpm)were obviously increased in model group(all P<0.05). Compared with the model group,the LVSP,+dp/dt max,-dp/dt max, (dp/dt)?P-1max, HMI,LVMI were significantly increased in myocardial injection group and acupoint injection group〔LVSP:130.38±14.96,124.36±14.36 vs. 114.36±12.71,+dp/dt max:4707.52±394.36,4597.14±411.05 vs. 3791.43±327.29,-dp/dt max:4075.11±317.89,3938.05±373.76 vs. 3116.32±275.04,(dp/dt)?P-1max:215.26±21.29,197.39±18.96 vs. 155.93±25.14〕,and the LVEDP and HR were significantly decreased(LVEDP:5.15±2.39,5.64±1.96 vs. 10.58±2.49,HR:400.50±42.58,395.55±44.62 vs. 414.51±35.75,all P<0.05). There were no significant differences in above indexes between myocardium injection group and acupoint injection group. Conclusion Acupoint injection of BMSCs can improve the heart function of rat model with AMI.
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Objective To investigate the effect of intermittent tensile strain on the proliferation and osteogenic differentiation of rBMSCs (rat bone mesenchymal stem cells). Methods Intermittent tensile strain was applied on rBMSCs in vitro by Flexcell 4 000 Tension System (10% elongation amplitude, 0.5 Hz, twice every day, 4 h every time), then effects of the strain after 1, 3, 5, 7 d on cell morphology, cell proliferation, and the relative expression of Cbfα1（core binding factor α1），ALP and collagen I mRNA as well as Cbfα1 protein were measured. Results Intermittent tensile strain slowed the proliferation of rBMSCs from the first day to the seventh day. The relative expression of ALP and collagen I mRNA increased by 3~6 times from the third day（P＜0.05）, meanwhile the expression of Cbfα1 mRNA and protein was up-regulated under the mechanical stimulation. Conclusions Mechanical stretch plays an important role in the proliferation and differentiation of rBMSC, and approprite intermittent tensile strain can slow the proliferation of rBMSC and promot its osteogenic differentiation.
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ObjectiveTo investigate the role of p38 and ERK1/2 during rhesus monkeys mesenchymal stem cells differentiated into neuron-like cells.MethodsTo induce the neuronal phenotype,rhesus monkeys mesenchymal stem cells were maintained in sub-confluent cultures in serum-contain medium supplement with Sonic hedgehog.Western blot analysis the change of p38 and ERK1/2 during rhesus monkeys mesenchymal stem cells differentiated into neuron-like cells.Under transmission and scanning electron microscope,ultra-structure of the differentiated cell were observed.ResultsDuring BMSCs differentiated into neuron-like cells by SHH,Mitogen-activated protein kinases (MAPK) involved in their signal transduction,p38 was activated and ERK1/2 was inhibited.P38 inhibitor SB203580 increased induced differentiation time compared with normal induced cells,and inhibited neurite outgrowth.ConclusionActivation of p38 and inhibition of ERK was impacted on differentiation into neuron-like cells from rhesus monkeys mesenchymal stem cells induced by Sonic hedgehog,which may has potential application on neuroprotection of stem cells in Nervous system diseases
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BackgroundLimbal stem cell deficiency usually leads to blindness, and traditional therapy is limited. Recent research demonstrated that bone mesenchymal stem cells ( BMSCs ) and human amniotic epithelial cells(AECs) could differentiate into many kinds of cells including corneal epithelial cells, but the outcome and effect of these cells on corneal stem cell deficiency are still unclear. ObjectiveThis study aimed to observe and compare the effects of rabbit BMSCs and human AECs transplantation for rabbit limbal stem cell deficiency. Methods Eighteen clean New Zealand rabbits were randomly divided into the amniotic stroma(AS) group, rabbit BMSCs group and human AECs group with 6 rabbits for each group. Limbal stem cell deficiency models were established by putting a piece of filter paper that had been soaked in a NaOH solution at the corneal center. Rabbit BMSCs were isolated and purified by density gradient centrifugation combined with the attachment culture method, and human AECs were collected by a sequential trypsin digestion technique,and the third generation rabbit BMSCs and the first generation human AECs were identified with RT-PCR. After that,cells were inoculated onto the denuded AS and grafted to the corneal surface of the experimental animals. Twenty-eight days after cell transplantation, the therapeutic effects were evaluated based on the corneal neovascularization and opacity scores. Corneal histopathological examination and immunohistochemistry were performed to evaluate and compare the effectiveness among AS,rabbit BMSCs and human AECs on corneal stem cell deficiency. The procedure complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. ResultsThe third generation of rabbit BMSCs grew well after 12 hours, and the first generation of human AECs formed a membrane-like monolayer after 48 hours of incubation on AS. Immunohistochemistry staining showed that, 28 days after transplantation, the surface cells of the cornea were positive for cytokeratin 3 in both the rabbit BMSCs group and human AECs group.Compared with the AS group,the corneal neovascularization and opacity grades were significantly decreased in the rabbit BMSCs group( Z=-2. 983, P =0. 003 ; Z =-2. 844, P =0. 004 ) and human AECs group ( Z =-2. 817, P =0. 005 ; Z =-2.041, P =0. 041 ). Histopathological analysis exhibited that stratified corneal epithelial-like cells formed on the corneal stroms 28 days after grafting and no signs of goblet cells and neovascularization were found. Less inflammatory cells and regular collagen fiber could be seen in the rabbit BMSCs group and human AECs group. In addition,clinical observation also revealed that the corneas were much clearer in the rabbit BMSCs group than the human A ECs group( Z =-2. 091 , P=0. 037 ), but the corneal neovascularization score was similar between them (Z = -0. 267,P=-0. 789). ConclusionsRabbit BMSCs and human AECs can differentiate into corneal epithelial-like cells onthedamagedcornealsurfaceandfurtherdemonstrateremarkableinhibitoryeffectsoncornealneovascularization and inflammatory cells. The more dominant and prominent effect is the role played by rabbit BMSCsin the improvement of corneal transparency.
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The rats were assigned to blank control group, classical induction group, and drynaria total flavonoid group. Whole bone marrow culture method was applied to isolate and purify rats bone mesenchymal stem cells (BMSCs). Akaline phosphatase activity, calcium nodes, TGF-β1 and BMP-2 secretion in the process of bone mesenchymal stem cells osteogenic differentiation were detected. The results showed that compared to the blank group and classical group, drynaria total flavonoid promoted osteogenic differentiation accompanied with increased TGF-β1 and BMP-2 secretion (all P＜0. 05). Drynaria total flavonoid may promote osteogenic differentiation of BMSCs via upregulating TGF-β1 and BMP-2 expressions, and play an active role in the treatment of osteoporosis.
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Objective To induce bone marrow mesenchymal stem cells(BMSCs) or bone marrow stroma stem cell(MSCs) to differentiate directionally towards chondrocytes in vitro and then identify the differentiated cells.Methods Bone marrow was harvested from the iliac bone of 16-week-old Japanese white rabbits.After gradient centrifugation,cultivation,amplification,the 3rd-passaged BMSC were implanted in six-hole-plate according to a certain proportion and induced by chondrogenic inducers including transforming growth factor-?1,dexamethasone and vitamin C.Chondrocytes were selected and fixed at different time.The features of chondrocytes were identified by toluidine blue staining and collagen types II immunohistochemical assay.Results The structure of cellular cartilage from BMSCs was uniformly positive of toluidine blue staining and collagen types II immunohistochemical staining.Conclusion Rabbit BMSC,obtained under the experimental conditions,develops stably,proliferates rapidly and is differentiated successfully into chondrocytes by induction in vitro.
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Objective To observe the protective and repairing effects of bone mesenchymal stem cells(BMSCs) transplantation on cerebral infarction in rats and to study the different effect of transplantation at different time points. Methods Middle cerebral artery occlusion(MCAO) models were set up,and the rats were divided into a control group,a group with PBS transplantation and five groups with BMSCs transplantation 3,6,12,24 and 72 h after MCAO,respectively.The volume of infarction area and the neurological severity score(NSS) in all the groups were compared. Results Twenty-eight days after MCAO,the TTC staining indicated that the volume of infarction area in the groups with BMSCs transplantation decreased remarkably compared with the control group and the group with PBS transplantation(P
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@#ObjectiveTo investigate the feasibility of construction transplantable tissue engineered bone in nude mouse as in vivo bioreactor. MethodsSelected the 4th passage of rabbit bone mesenchymal stem cells, which were cultured, expanded and induced in vitro. Cells were seeded into porous natural coral. The complexes were implanted subcutaneous in nude mice. Regenerated bone was taken out 8 weeks after implantation and implanted into muscle tissue interspace in the back of rabbit itself.The rabbit-anti-mouse antibody titer was measured at 14,28,56 days and the level of IL-2 was detected the at 2, 7, 14, 28 and 56 days after implantation.Immunohistochemistry staining was carried on in the 8th week. ResultsDuring the whole experimental process, IL-2 activity couldn't be measured.The rabbit-anti-mouse antibody titer was positive.A specific antibody reaction was detected from 2ed week to 8th week after transplantation and touches the peak value four weeks later.Immunohistochemistry staining result was negative.ConclusionThe tissue engineered bone growth in nude mouse is seemed to be the reliable transplantable bone source.
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@# ObjectiveTo evaluate the effectiveness of mesenchymal stem cell (MSC) in osteonecrosis of the femoral head defect(ONFH) repair. MethodsAniaml model of ONFH defect were established with rabbits, which were divided into 3 groups,group A did not infill anything as control, nano Hydroxyapatite/collagen(nHAC) as group B,nHAC+MSC as group C.Histology change were investigated 4,8,12 weeks after operation respectively.Results groups B and C were different with group A. The difference between groups A and C was more significant. ConclusionMesenchymal stem cell has a strong activity of osteoconduction,it has a value in repairing the bone defect of ONFH and the treatment of ONFH.
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Objective:To explore the isolation,culture,proliferation and fluor-labelling of the fetal bone mesenchymal stem cell (BMSCs),and their biological character. Methods:Limb bone marrow samples from fetus aged 4 to 5 months were obtained, BMSCs were isolated by Ficoll centrifugalization and adherence screening method,and then cultured,proliferated to the 12th generation in vitro. The growth curve,cell cycle,phenotype,enzymatic activity,glycogen and the DAPI fluor-labelling of BMSCs were observed. Results:(1)Major BMSCs were fusiform, arranged as whirlpool. After the 12th generation,BMSCs presented the signs of senility. The subcultured cells proliferated faster than primary cells. (2)The 3rd and 8th generation of BMSCs expressed CD29,D71,but not CD34,HLA-DR;the G0,G1 period cells were 93.73%,and the S,M,G2 period cells were 6.27%. (3)The 5th generation cells were positive of nonspecific esterase and glycogen,but weak positive of alkaline phosphatase. (4)DAPI labeled BMSCs grew normaly,and the marking rate of BMSCs was 100%,but decreased gradually,and only 15% cells were labbelled for the 3th week. Conclusion:The acquired fetal BMSCs have the character of MSCs. The DAPI labeling is a modus operandi for the short period tracing BMSCs.
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Objective: To examine the effects of alginate on bone mesenchymal stem cells(bMSCs).Methods: bMSCs were cultured on the alginate gels(experimental group) and tissue culture plastics(control group);the morphous of bMSCs was observed by microscope after toluidine blue staining;the growth curves were analyzed by MTT;and the expressions of osteoblast related genes(alkaline phosphatase,collagen I and osteonectin) were detected by RT-PCR when cultured for 6 days.Results: There were no difference in the cell proliferation between the experimental group and control group.The morphous of bMSCs in the experimental group was induced to be similar to that of osteoblasts and form colony proliferation obviously.The alkaline phosphatase and collagen I genes were positive in the two groups,but their expressions in the experimental group were higher;at the same time,osteonectin gene was positive in the experimental group and negative in the control group.Conclusion: bMSCs may differentiate into osteoblasts automatically when cultured for longer time and that alginate gels could promote this differentiation and may be suitable biomaterial of scaffold in bone tissue engineering.
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Objective To explore the distribution and differentiation of bone mesenchymal stem cells (MSCs) in the brain of neonatal rats with hypoxic ischemic encephalopathy (HIE). Methods MSCs were isolated and purified by adhering to the culture glassware wall and prelabeled with bromodeoxyuridine (BrdU) for 72 h before transplantation. The model of HIE was established. At 24 h after hypoxic ischemia, approximate 4?10 6 cells were injected into the brain of neonatal rats with HIE through the right side bregma. The Nestin, neuron specific enolase (NSE) and glial fibrillary acid protein (GFAP) were detected by immunofluorochemistry at 4 weeks after injection. Results Majority of MSCs were distributed in the cortex, hippocampus of the lesioned hemisphere. The number of MSCs was (2781?254) in the left hemisphere, but (4708?281) in the right hemisphere. There was significant difference (t=18.70, P0.05). The expression ratio of NSE was (3.79?0.95)% in the left hemisphere, but (5.69?1.48)% in the right hemisphere (t=3.404, P0.05). Conclusion MSCs are mainly distributed in the lesioned hemisphere and can differentiate into neuronal-like cells, express the mark of neural stem cells, neurons and neuroglial cells at 28 d after intracortical transplantation.