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Aim To investigate the effect of quercetin on the aging model of bone marrow mesenchymal stem cells established under microgravity. Methods Using 3D gyroscope, a aging model of bone marrow mesenchymal stem cells was constructed, and after receiving quercetin and microgravity treatment, the anti-aging effect of the quercetin was evaluated by detecting related proteins and oxidation indexes. Results Compared to the control group, the expressions of age-related proteins p21, pi6, p53 and RB in the microgravity group significantly increased, while the expressions of cyclin D1 and lamin B1 significantly decreased, with statistical significance (P<0.05). In the microgravity group, mitochondrial membrane potential significantly decreased (P<0.05), ROS accumulation significantly increased (P <0.05), SOD content significantly decreased and MDA content significantly increased (P<0.05). Compared to the microgravity group, the expressions of age-related proteins p21, pi6, p53 and RB in the quercetin group significantly decreased, while the expressions of cyclin D1 and lamin B1 significantly increased, with statistical significance (P<0.05). In the quercetin group, mitochondrial membrane potential significantly increased (P<0.05), ROS accumulation significantly decreased (P<0.05), SOD content significantly increased and MDA content significantly decreased (P<0.05). Conclusions Quercetin can resist oxidation, protect mitochondrial function and normal cell cycle, thus delaying the aging of bone marrow mesenchymal stem cells induced by microgravity.
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Macrophages are typically identified as classically activated (M1) macrophages and alternatively activated (M2) macrophages, which respectively exhibit pro- and anti-inflammatory phenotypes, and the balance between these two subtypes plays a critical role in the regulation of tissue inflammation, injury, and repair processes. Recent studies indicate that tissue cells and macrophages interact
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With the development of the 3rd-generation high-throughput sequencing technology and tissue engineering, recent studies show that many long-chain non-coding RNAs (LncRNAs) have played an important role in osteogenic differentiation of mesenchymal stem cells (MSCs). LncRNAs, which are involved in the regulation of mechanical regulation, further regulate bone-related cell functions and play a regulatory role at multiple levels, including transcription, post-transcriptional and epigenetic. LncRNAs may be involved in the osteogenic differentiation and bone remodeling of MSCs, the regulation of bone-related cell functions as a mechanical response molecule, as well as the pathological process of skeletal diseases.
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Objective To investigate the effect of interleukin( IL)-6 secreted by bone marrow stromal cells HS-5 on activity and apoptosis of human acute myeloid leukemia( AML) cells HL-60 and its possible mechanism. Methods HL-60 cells and HS-5 cells were cultured in vitro, and the co-culture system was established. Scanning electron microscope, ELISA, cell counting kit-8( CCK-8) , AnnexinV-FITC/PI, double-staining flow cytometry, Real-time PCR and Western blotting techniques were used to detect the changes of viability and apoptosis of HL-60 cells, respectively. HL-60 cells from different groups were inoculated into BALB/c nude mice to observe and record tumorigenesis. Results IL-6 secreted by bone marrow stromal cells HS-5 could enhance the viability and inhibit the apoptosis of HL-60 cells, and down-regulate the expression of pro-apoptotic gene Bax and and up-regulate the expression of anti-apoptotic gene Bcl-2 in HL-60 cells. HL-60 cells in co-culture group had the strongest tumorigenicity in BALB/c nude mice, while HL-60 cells alone had the weakest tumorigenicity. Conclusion Part of the mechanism by which bone marrow stromal cells HS-5 promote the proliferation of HL-60 cells and inhibit their apoptosis may be through the secretion of IL-6.
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Objective @#To investigate the inductive effects of canine periodontal ligament stem cells (PDLSCs) cocultured with canine disparate differentiating-period iliac bone marrow stromal cells (I-BMSCs).@*Methods@#Purified PDLSCs were isolated by in vitro culture of cBMSCs and flow cytometry. Third-generation PDLSCs were obtained, and conditioned culture medium derived fromiliac bone marrow stromal cells (I-BMSCs-CM) was added as indicated for coculture of PDLSCs. As the control group, pure uninduced PDLSCs were routinely cultured in DMEM culture medium containing 15%FBS. The experimental groups included the I-BMSCs-CM, I-BMSCs-CM-10ds and I-BMSCs-CM-15ds groups. The I-BMSCs-CM group consisted of PDLSCs induced by I-BMSCs, the I-BMSCs-CM-10ds group consisted of PDLSCs induced by I-BMSCs after osteogenic induction for 10 days, and the I-BMSCs-CM-15ds group consisted of PDLSCs induced by I-BMSCs after osteogenic induction for 15 days. The cocultured PDLSCs were examined via the MTT assay. Total mRNA and protein were prepared at 3 and 7 days. The mRNA expression levels of runt-related transcription factor 2(Runx2), special AT-rich sequence binding protein 2(Satb2) and osteocalcin (OCN) were measured by qRT-PCR. The protein expression levels of Satb2, Runx2 and OCN were detected by Western blot.@*Results@#The PDLSCs showed a spindle-like morphology. While the BMSC-conditioned media increased PDLSCs proliferation, the media conditioned by BMSCs allowed to differentiate for 15 days (I-BMSCs-CM-15days) significantly enhanced PDLSCs proliferation (F=342.8, P=0.017). The expression levels of the analyzed genes were upregulated in the coculture groups, and the protein expression levels of Satb2, Runx2 and OCN were higher in the test groups than in the control group at 7 days. At the protein level, I-BMSCs-CM-15days upregulated the expression of Satb2 by 3.04-fold (FSatb2=24.48, P=0.014), Runx2 by 5.1-fold (FRunx2=12.25, P < 0.001), and OCN by 3.67-fold (FOCN=18.35, P=0.022).@*Conclusion @#The conditioned medium of I-BMSCs may enhance the proliferation of PDLSCs, and that of terminally differentiated bone cells probably triggered the osteogenesis of PDLSCs, suggesting important implications for periodontal engineering.
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ObjectiveTripartite motif 33 (Trim33) is known to play a very important part in regulating osteoblast differentiation, but its role in adipocyte differentiation is rarely reported. The aim of this study was to investigate the regulatory effect of Trim33 on adipocyte differentiation.MethodsBone marrow stromal cell ST2 cells transfected with the Trim33-pcDNA3.1 plasmid were included in the experimental group and those transfected with the pcDNA3.1 plasmid taken as the control. The cells of both groups were treated with adipogenic medium to induce adipocyte differentiation, followed by determination of the expressions of the adipocyte-specific genes CCAAT enhancer binding protein alpha (C/EBPα), peroxisome proliferator-activated receptor gamma (PPARγ), adipocyte characterizing factor FABP4, and adipocytokines adipsin by oil red O staining, qRT-PCR and Western blotting.ResultsThe expression of Trim33 was significantly upregulated in the experimental group as compared with that in the control (88.51±14.31 vs 1.00±0.31, P<0.01). After 5 days of adipogenesis induction, there were dramatically more lipid droplets in the ST2 cells and the A value was markedly higher in the former than in the latter group (0.69±0.03 vs 0.34±0.03, P<0.01). Compared with the control, the cells in the experimental group exhibited remarkable increases in the relative mRNA expressions of C/EBPα, PPARγ, FABP4 and adipsin as well as the protein expressions of Trim33, PPARγ, C/EBPα and FABP4.ConclusionTrim33 promotes lipid accumulation and upregulates the expressions of adipocyte-specific genes in bone marrow stromal cells.
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Objective To observe the effect of lentiviral vector-mediated basic fibroblast growth factor (bFGF) gene transfection on the biological characteristics of rabbit bone marrow stromal cells(BMSCs)under in vitro culture conditions. Methods BMSCs were obtained by density gradient centrifugation and adherence screening. The bFGF gene was transfected into BMSCs by lentiviral vector and divided into bFGF transfection group,empty virus group and untransfected group according to the transfection conditions.After transfection,the morphology,expressions of bFGF mRNA and protein, cell proliferation,cell cycle and alkaline phosphatase(ALP)activity were observed in three groups of cells. Results High density BMSCs were successfully obtained by density gradient centrifugation and adherence screening.After transfection of BMSCs with bFGF gene, the cell morphology showed no significant changes, while the expressions of bFGF mRNA and protein were significantly increased, the cell proliferation curve shifted upward, the proportion of proliferating cells increased,and the activity of ALP was significantly enhanced.There were significant differences between three groups(P<0.05).Conclusion The rabbit bFGF gene is successfully introduced into the BMSCs cultured in vitro by lentiviral vector, and the target gene is stably expressed.The expression of bFGF can promote the proliferation and osteogenic differentiation of BMSCs.
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Objective To construct miR-196b sponge lentiviral vector,and laid the foundation for studying the function of miR-196b in bone marrow stromal cells.Methods Based on the miR-196b mature sequence,a sequence consisting of 6 tandem repeats of the complementary sequence of miR-196b was designed,and which was cloned into pUC19 plasmid by using reverse PCR.Then the six-repeat sequence was cut and subcloned into pLVX-shRNA2 lentiviral vector.The lentivirus was packaged using 293T cells,and titer determination was done.The pLVX-shRNA2 lentivirus was used as the control group,and the 196b-sponge-pLVX lentivirus was the experimental group.Then ST2 cells were infected with the viruses,and the infection efficiency was calculated.The protein level of forkhead box O1 (FoxO1) was detected by Western blot assay.Results The identity of the sponge sequence was verified by sequencing.The titer of the sponge virus was 1 × 108 PFU/mL,and the infection efficiency reached 80％.Compared with the control group,the expression level of FoxO 1 protein was significantly increased (P ＜ 0.05).Conclusion The miR-196b sponge lentiviral vector is successfully constructed,and which has the capability to inhibit endogenous miR-196b.
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Objective To investigate the effects of human leptin ( hLEP) gene transfection on rat bone marrow stro-mal cells ( rBMSCs) .Methods rBMSCs were cultured and transfected with adenoviruses encoding hLEP ( Ad5-hLEP-EGFP) in vitro as experimental group while rBMSCs transfected with Ad 5-EGFP and non-transfected were control groups.The proliferation was detected by MTT and the expression of collagen type Ⅰ(Col-Ⅰ) and alkaline phosphatase ( ALP) were assessed by real-time PCR.The ability of mineralized nodule forming was also examined by Alizarin red staining .The combination of transfected rBMSCs and β-tricalcium phosphate (β-TCP ) was con-structed and the osteogenic ability of the construction was evaluated in nude mice .Results hLEP could be trans-fected into rBMSCs successfully by adenovirous .After transfection , the proliferation was not affected while Col-Ⅰand ALP expressions were more pronounced in rBMSCs transfected with Ad 5-hLEP-EGFP ( P<0.05 ) .Alizarin red staining showed the ability of mineralized nodule forming was also up-regulated in Ad5-hLEP-EGFP group (P<0.05).In addition, the transfected rBMSCs adhered to β-TCP and survived well and the combination showed more new bone like tissue formation in nude mice compared to control groups .Conclusions rBMSCs transfected with hLEP might be potently used in bone or periodontal tissue regeneration .
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To investigate the effect of icaritin (ICT) combined with GDF-5 on chondrogenic differentiation of bone marrow stromal cells (BMSCs), and discuss the action of Wnt signaling pathway, full bone marrow adherent method was used to isolate and culture SD rats BMSCs, and the cells at P3 generation were taken and divided into 6 groups： BMSCs group, ICT group, GDF-5 group, GDF-5+ICT group, GDF-5+ICT+SB216763 group, and GDF-5+ICT+ XAV-939 group. The cells were induced and cultured for 14 days. The morphology change was observed by inverted microscope. Alcian blue staining method was used to detect the changes of proteoglycans. RT-PCR was used to detect the mRNA expressions of aggrecan, Col2, Sox9, Dvl1, Gsk3β, and β-catenin. The protein expressions of collagen 2 (COL2) and β-catenin were detected by Western blot. The results indicated that, compared with the BMSCs group, gradual increase was present in proteoglycan Alcian blue staining; mRNA expressions of cartilage differentiation marker genes aggrecan, COL2, Sox9 and the protein expression of COL2, as well as mRNA and protein expressions of Wnt signaling pathway-related gene β-catenin, but with gradual decrease in Gsk3β mRNA expressions in GDF-5 group, GDF-5+ICT group and GDF-5+ICT+SB216763 group. On the contrary, compared with GDF-5+ICT group, there was a decrease in expressions of Dvl1, and β-catenin related to chondrogenic differentiation and Wnt signaling pathway, a increase in Gsk3β mRNA expression, and also a decrease in protein expressions of COL2 and β-catenin in GDF-5+ICT+XAV-939 group, with statistically significant difference between two groups. GDF-5 in combination with icaritin can induce chondrogenic differentiation of BMSCs in rats, and icaritin (ICT) can promote the chondrogenic differentiation. ICT can promote the chondrogenic differentiation of BMSCs in vitro probably by activating the Wnt/β-catenin signaling pathway.
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Objective To investigate the effects of bone marrow stromal cells(BMSCs)transplantation combined with low dose ultrashort wave (USW)radiation on functional recovery and the expression of glial fibrillary acidic protein(GFAP)and ED?1 after spinal cord injury(SCI)in rats,and further discuss its action mechanism. Methods Female Sprague?Dawley rats(n=30)were randomly divided into 5 groups:sham?oper?ated,as well as control,USW,BMSCs,and USW+BMSCs that were subjected to spinal cord injury(SCI). Basso?Beattie?Bresnahan(BBB)tests were carried out before the operation and at 1 d,1 week,2 weeks,3 weeks,4 weeks after SCI. 4 weeks later,animals were sacrificed and tissues were collected to make paraffin section. Immunohistochemical staining was performed to observe the expression of GFAP and ED?1. Results 4 weeks after SCI,BBB scores were significantly higher in the USW and USW+BMSCs groups than in the control group(both P<0.001). No signifi?cant difference was observed between the BMSCs group and control group. On the expression of GFAP ,only USW+BMSCs group showed signifi?cantly decreased compared with the control group(P<0.05). All treatment groups exhibited lower ED?1 expression than the control group(all P<0.05). Conclusion Our results indicate that USW radiation alone can obviously improve neural functional recovery after SCI. The USW radi?ation and BMSCs transplantation treatment can reduce inflammation ,and USW radiation is more effective. The combination therapy did not show a synergistic action on promoting functional recovery ,but do have an effect on reducing the inflammatory response and glial scar formation.
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Objective To investigate the possibility of the domestic reticulated vitreous carbon as a kind of scaffold material for bone tissue engineering,the biocompatibility of domestic reticulated vitreous carbon was first successfully tested with bone marrow stromal cells (BMSCs) in vitro and for bone tissue repair in vivo.Methods From June,2013 to August,2014,the morphology and proliferation of BMSCs co-cultured with scaffold material in vitro was measured.Differences of measurement were compared with single factor analysis of variance to detect the cytotoxicity of reticulated vitreous carbon.In vivo reticulated vitreous carbon were implanted into the bone defect site and the groin.After 12 weeks,the biocompatibility of reticulated vitreous carbon was observed.Results MTT results showed that after 7d co-culture,the survival and proliferation of BMSCs had not been significantly inhibited (P ＞ 0.05).Inverted fluorescence microscope and scanning electron microscope found that newly developed three-dimensional domestic reticulated vitreous carbon could promote adhesion,aggregation and proliferation of BMSCs in vitro.Studies in vivo demonstrate that implanted reticulated vitreous carbon with a high porosity and host bone may produce a stable connection and integration.Conclusion Non-cytotoxic domestic reticulated vitreous carbon can promote the adhesion and proliferation of bone marrow mesenchymal stem cells in vitro and has good bone induction properties in vivo.
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Objective To investigate the effect of geraniin on expression of Wnt3a protein and mRNA in bone marrow stromal cell (BMSC) from osteoporotic rats. Methods The model of osteoporosis (OP) was duplicated by ovariectomy in rats. BMSCs were isolated and cultured. BMSCs from shamed rats were routinly cultured and taken as normal control, and BMSCs from OP rats were divided into model group, 1μmol/L simvastatin positive group, and geraniin group (0.01, 0.1, 1, 10, 100 μmol/L), respectively. The methods of realtime-PCR and western blot were used to assay the protein and mRNA expression of wnt3a, respectively.Results As compared with normal control group, the protein and mRNA expression of wnt3a in model group were significantly suppressed;Compared with model group, 1 μmol/L simvastatin, and 0.1, 1, 10 and 100 μmol/L geraniin significantly increased the expression of wnt3a protein and mRNA. Conclusion It is suggested that geraniin activates wnt/β-catenin pathway though increasing the expression of signaling protein wnt 3a in BMSCs from OP rats. It would be beneficial to osteogenic differentiation of BMSCs and osteogenesis.
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BACKGROUND: Bortezomib is widely used for the treatment of multiple myeloma. Bone marrow stromal cells (BMSCs) endow myeloma cells with survival and growth advantages. However, the influence of bortezomib on BMSCs is not well elucidated. We examined the effects of bortezomib on the survival and growth of BMSCs in vitro. METHODS: The effects of bortezomib on the survival and proliferation of the BMSC MS-5 cell line and on BMSCs obtained from healthy individuals (N=4) and newly diagnosed myeloma patients (N=5) were investigated in vitro. Transmembrane cell migration was evaluated using the Transwell system. A short interfering RNA strategy was used to knock down the expression of chemokine (CXC motif) ligand 12 (CXCL12) mRNA. To examine the effects of bortezomib-exposed BMSCs on the migration and localization of myeloma cells, MS-5 monolayers were treated with bortezomib for 24 hr, washed, and then overlaid with human RPMI8226 myeloma cells. RESULTS: Bortezomib inhibited BMSC proliferation in a concentration-dependent manner, and induced cellular apoptosis. Bortezomib decreased CXCL12 production by BMSCs. Knockdown of CXCL12 mRNA in BMSCs revealed that CXCL12 served as an autocrine growth factor. Short-term bortezomib treatment of BMSC monolayers reduced the tendency of myeloma cells to locate to positions under the monolayers. CONCLUSION: Bortezomib inhibits the survival and growth of BMSCs via downregulation of CXCL12, which may contribute to the clinical effects of this agent.
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Humans , Apoptosis , Cell Line , Cell Movement , Down-Regulation , Mesenchymal Stem Cells , Multiple Myeloma , RNA, Messenger , RNA, Small Interfering , BortezomibABSTRACT
Objective The effect of Asperosaponin Ⅵ(ASAⅥ)on adipocyte differentiation and the involvement of Wnt signal pathway was investigated. Methods The murine bone marrow stromal cell line ST-2 were divided into 6 groups:control group, adipocyte differentiation group, and 4 different doses of ASAⅥgroups. Control group was exposed to the vehicle, adipocyte differentiation group was exposed to adipogenic reagent, and those 4 ASAⅥgroups were treated with different concentration(10-7, 10-6, 10-5, 10-4 mol/L)of ASAⅥafter adipocyte differentiation induction. 5 days later, oil red O staining was performed to calculate adipocyte rate. Then mRNA transcription levels of PPARγ, FABP4 genes andβ-catenin that were Wnt/β-catenin signaling pathway proteins were examined by FQ-PCR. Then Wnt pathway inhibitor DKK1 was supplemented into ST-2 cells treated with 10-4 mol/L ASAⅥfor 5 days. After that FQ-PCR was used to detect whether tran?scription levels of PPARγ, FABP4 andβ-catenin in ST-2 cells were changed. Results Compared with adipocyte differenti?ation group 10-5 mol/L and 10-4 mol/L ASAⅥtreatments greatly down-regulated the number of lipid droplets and markedly inhibited transcription levels of adipocyte characterization transcription factors included PPARγ, FABP4 while up-regulat?ed transcription level ofβ-catenin in ST-2 cells. DKK1 can reverse the inhibitory effect of ASAⅥon adipocyte differentia?tion in ST-2 adipocyte. The transcription levels of PPARγand FABP4 were up-regulated significantly while transcription level ofβ-catenin was inhibited. Conclusion ASAⅥblocks adipocyte differentiation in ST-2 cells which might be medi?ated through activating Wnt/β-catenin signaling pathway.
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BACKGROUND: Sevoflurane exposure during the early postnatal period causes neuroinflammation and neuronal apoptosis in rodents. Bone marrow stromal cells (BMSCs) have been shown to protect and repair the damaged central nervous system, for example in ischemic stroke models. In this study, we investigated whether intravenous administration of BMSCs ameliorated neurodegeneration, induced by sevoflurane exposure, in neonatal rats. METHODS: Sprague-Dawley rat pups (postnatal day 7) were exposed to 2% sevoflurane for 6 h (vehicle group, n = 7). BMSCs were administered 30 min after induction of sevoflurane anesthesia (BMSCs group, n = 7). The pups were exposed to carrier gas only, as a negative control (mock anesthesia group, n = 4). We assessed the therapeutic effects of BMSC treatment by measuring expression of the pro-inflammatory cytokine interleukin-6 (IL-6), and levels of cleaved caspase-3, in brain tissues immediately following sevoflurane anesthesia. RESULTS: Analysis of the cleaved caspase-3 bands revealed that levels of activated caspase-3 were elevated in the vehicle group compared with the mock anesthesia group, indicating that a single exposure to sevoflurane at subclinical concentrations can precipitate neuronal apoptosis. BMSC treatment did not suppress apoptosis induced by sevoflurane exposure (compared with the vehicle group). The vehicle group had higher proinflammatory cytokine IL-6 protein levels compared with the mock anesthesia group, indicating that sevoflurane exposure induces IL-6 expression. BMSC treatment suppressed sevoflurane-induced increases in IL-6 expression, indicating that these cells can inhibit the neuroinflammation induced by sevoflurane exposure (vehicle group vs. BMSC group). CONCLUSIONS: Intravenous administration of BMSCs reduces neuroinflammation, but does not attenuate apoptosis induced by sevoflurane exposure.
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Animals , Rats , Administration, Intravenous , Anesthesia , Apoptosis , Bone Marrow , Brain , Caspase 3 , Central Nervous System , Interleukin-6 , Mesenchymal Stem Cells , Neurons , Rats, Sprague-Dawley , Rodentia , StrokeABSTRACT
Primary human bone marrow stromal cells (hMSCs) were transfected with human telomerase reverse transcriptase (hTERT) gene with lipofection method. The hTERT transfected hMSCs of passage 100 underwent chondrogenesis induction with dexamethasone, transforming the growth factor β and vitamin C, osteogenesis induction with dexamethasone, β glycerophosphoric acid and vitamin C, and cardiomyocyte induction with 5-azacytidine. After 7, 14, 21 and 28 days of induction, immunocytochemistry was performed to detect the expressions of type I and II collagen and osteocalcin, and alizarin red staining was performed to detect the bone nodule formation in osteogenesis induction. Immunocytochemistry was carried out to detect the striated muscle actin expression in cardiomyocytes. The hMSCs undergoing successful transfection were positive for the hTERT. The hTERT transfected cells were grown in vitro successfully and passaged for 136 generations. Results showed that these cells could be induced to differentiate into chondrocytes, bone and myocardial cells. Introduction of exogenous hTERT into hMSCs could achieve immortalized hMSCs with the potential of multi-directional differentiation. Thus, these cells could be applied as seed cells in tissue engineering.
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Objective To evaluate differences in genes expression of rat bone marrow stromal cells (rBMSCs) under continuous mechanical strain by gene microarray technology．Methods rBMSCs were isolated and cultured in vitro. Continuous stresses with amplitude of 10% and frequency of 1 Hz were applied on rBMSCs for 6 hours by Flexercell mechanical loading system to investigate rBMSC gene expression profiles, and quantitative PCR was used to verify gene expression changes related to osteoblastic differentiation. Results Compared with the control group, 1 244 differentially expressed genes were found in mechanical loading group, among which 793 genes were up-regulated, while 451 genes were down-regulated．GO (gene ontology) analysis suggested that differentially expressed genes were mainly involved in multicellular organismal development, cell differentiation, chemotaxis, cell adhesion and so on. Four signaling pathways as Notch, Wnt, FGF and IGF might participate in the regulation of stress-induced osteoblastic differentiation. PCR validation results were consistent with the gene chip results. Conclusions Mechanical stress could induce osteoblastic differentiation of the BMSCs, while several differentially expressed genes screened by gene microarray may attribute to this process.
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OBJECTIVE@#To study the changes of gene expression profiles associated with osteoblasts differentiated from rat bone marrow stromal cells in vitro by gene chip technique.@*METHODS@#rat Bone marrow stromal cells were isolated and cultured, and differentiation was induced by dexamethasone, β-glycerol phosphate and vitamin C. Cellular mRNA was extracted and reverse transcribed into cDNA, thus related genes expression differences were detected by gene expression profile chip.@*RESULTS@#Calcifying nodules were visible in the induced cells. There were 27.7% genes expressed differentially, three times more than the normal and induced cells, and some genes were related to transcription, translation, glycosylation modification. Extracellular matrix, signal molecules and metabolism were up-regulated.@*CONCLUSIONS@#The gene chip technique can be used to detect the multi-gene different expression in the differentiation-induced rat BMSCs, and these differentially expressed genes are necessary genes related to rat BMSCs proliferation and induction of osteoblastic differentiation.
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Animals , Rats , Cell Differentiation , Genetics , Physiology , Gene Expression Profiling , Mesenchymal Stem Cells , Cell Biology , Metabolism , Oligonucleotide Array Sequence Analysis , Osteoblasts , Cell Biology , Metabolism , Rats, Sprague-Dawley , Transcriptome , Genetics , PhysiologyABSTRACT
Objective To observe the synapse formation in hippocampal dentate gyrus, and the memory ability after transplanting bone marrow stromal cells (BMSCs) into lateral ventricle of ischemic brain injury rats. Methods BMSCs from femur of a Sprague-Dawley rat were cultured for 3 generations in vitro. 60 newborn Sprague-Dawley rats were divided into sham group (n=20), model group (n=20) and BMSCs group (n=20). The latter 2 groups were ligated the left common carotid artery, and the BMSCs group were injected BMSCs into the lateral ventricle 7 days after ligation. They were tested with Morris's water maze 8 weeks later, and then, the brains were immunohistochemi-cal staining to detect synaptophysin (SY). Results The time of probe trial of acquisition phase (T1) and reveral phase (T2) decreased in the model group compared with the sham group (P<0.05), and increased in the BMSCs group compared with the model group (P<0.05). The in-tegral optical density (IOD) of SY positive cells decreased in the model group compared with the sham group (P<0.05), and increased in the BMSCs group compared with the model group (P<0.05). Conclusion BMSCs implantation through lateral ventricle can improve the learn-ing and memory ability of rats, which may associated with the synapse formation in dentate gyrus.