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AIM: To investigate the potential of human induced pluripotent stem cells(hiPSCs)differentiating into corneal epithelial cells in the simulated limbal stem cells(LSCs)microenvironment.METHODS: The hiPSC cell lines were established in vitro, and hiPSCs were co-cultured with corneal stromal cells in transwell system, which simulated the LSC microenvironment. Bone morphogenetic protein 4(BMP4)and a specific transforming growth factor β inhibitor(SB431542)were added to improve the differentiation efficacy. The expression of corneal epithelial cell-specific markers CK3 and CK12, corneal epithelial cell precursor CK15, and the limbal stem cell markers ABCG5 were determined by immunofluorescence staining and flow cytometry.RESULTS: The hiPSCs were actively proliferated in vitro, and immunofluorescence staining showed positive stem cell-specific markers OCT4, SOX2, TRA-1-60 and NANOG. Furthermore, hiPSCs co-cultured with corneal stromal cells exhibited LSCs markers ABCG5 and corneal epithelial cell precursor markers CK15 were positive; however, corneal epithelial cell markers CK3 and CK12 were negative. With the addition of BMP4 and SB431542, hiPSCs showed positive expression of CK3, and the CK3 expression increased over the time.CONCLUSION: With the addition of SB431542 and BMP4, hiPSCs cultured in simulated LSCs microenvironment could differentiate into corneal epithelial cells.
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OBJECTIVE@#To explore the effect of dichloromethane extraction phase of ethanol extract from stem of Patrinia scabiosaefolia Fisch.(DPSS) on proliferation and differentiation of K562 cells and its related mechanism.@*METHODS@#MTT assay was used to detect the effects of DPSS at 0, 25, 50, 100 and 200 μg/ml on the proliferation of K562 cells at 24, 48 and 72 hours. Flow cytometry was used to analyze the changes of cell cycle and apoptosis at 24 and 48 hours. Wright-Giemsa staining was used to observe the morphological changes of K562 cells. The cell surface antigens CD33 and CD11b were detected by flow cytometry.@*RESULTS@#The proliferation of K562 cells treated with different concentrations of DPSS was inhibited in a time-dose dependent manner (r=-0.96). Cell cycle analysis showed that with the increase of DPSS concentration, cells in G2/M phase increased (r=0.88), and cells were blocked in G2/M phase. Flow cytometry results showed that with the apoptosis rate of K562 cells was the highest when treated with 200 μg/ml DPSS for 48 h. Morphological observation showed that the K562 cell body increased, the amount of cytoplasm increased, the ratio of nucleus to cytoplasm decreased, and the nuclear chromatin was rough after DPSS treatment. Cell differentiation antigen, CD33 and CD11b, were positively expressed after treated with DPSS.@*CONCLUSION@#DPSS can induce apoptosis through cell cycle arrest, inhibit the proliferation of K562 cells, and induce K562 cells to differentiate into monocytes, which has a potential anti-leukemia effect.
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Humans , K562 Cells , Patrinia , Methylene Chloride/pharmacology , Apoptosis , Cell Proliferation , Cell DifferentiationABSTRACT
The purpose of this study was to clone and characterize the ZFP36L1 (zinc finger protein 36-like 1) gene, clarify its expression characteristics, and elucidate its expression patterns in different tissues of goats. Samples of 15 tissues from Jianzhou big-eared goats, including heart, liver, spleen, lung and kidney were collected. Goat ZFP36L1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR), then the gene and protein sequence were analyzed by online tools. Quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression level of ZFP36L1 in intramuscular preadipocytes in different tissues and adipocytes of goat at different differentiation stages. The results showed that the length of ZFR36L1 gene was 1 224 bp, and the coding sequence (CDS) region was 1 017 bp, encoding 338 amino acids, which was a non-secretory unstable protein mainly located in nucleus and cytoplasm. Tissue expression profile showed that ZFP36L1 gene was expressed in all selected tissues. In visceral tissues, the small intestine showed the highest expression level (P < 0.01). In muscle tissue, the highest expression level was presented in longissimus dorsi muscle (P < 0.01), whereas the expression level in subcutaneous adipose tissue was significantly higher than that in other tissues (P < 0.01). The results of induced differentiation showed that the expression of this gene was up-regulated during adipogenic differentiation of intramuscular precursor adipocytes (P < 0.01). These data may help to clarify the biological function of the ZFP36L1 gene in goat.
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Animals , Goats/genetics , Amino Acid Sequence , Liver , Cloning, MolecularABSTRACT
【Objective】 To investigate the feasibility of differentiation of human AB plasma hematopoietic stem/progenitor cells (HSCs/HPCs) from peripheral blood into mature erythrocytes. 【Methods】 Hematopoietic stem/progenitor cells were induced to be differentiated into mature erythrocytes in the medium supplemented with 5% FBS, 3% FBS + 2% human AB plasma and 8% human AB plasma, respectively, and inoculated in 24-well culture plate at the density of 1×106/mL. Cell proliferation and morphological changes were observed in three different groups. Flow cytometry was used to detect erythroid terminal differentiation markers, i. e. GPA, Band3 and α4(α4-integrin), and late erythroid cell enucleation in different group. The effects of different culture conditions on HSCs/HPCs differentiation into mature erythrocytes were compared. 【Results】 The cell growth and proliferation multiples of the three groups (8% human AB plasma, 5% FBS and 3% FBS+ 2% human AB plasma) were 2 573±116 vs 2 514±246 vs 2 539±119(P>0.05), respectively. The morphological changes of the three groups were similar. With the extension of culture time, the cells differentiated from proerythroblasts to basophils, polychromatic erythroblasts and positive erythroblasts, and almost all of them differentiated into erythrocytes enucleation on day 21. GPA expression and enucleation rate(%) of the three groups were 97.17±1.91 vs 94.95±1.61 vs 96.15±1.38, and 85.1±3.26 vs 86.93±5.96 vs 86.5±3.36(P>0.05), respectively. 【Conclusion】 The differentiation of HSCs/HPCs from peripheral blood plasma into mature erythrocytes from human AB was similar to that of fetal bovine serum.
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@#In recent years, stem cell research and application in the field of ophthalmology has been highlighted, embryonic stem cells and adult stem cells can be targeted induced to differentiate into retinal pigment epithelial cells(RPE cells), which can obtain transdifferentiation of RPE cells source, through the body stem cells and retinal pigment epithelium cells transplantation is expected to be used in a variety of alternative treatment of the degenerative diseases of the retina. In this paper, the methods and application of various stem cells induced differentiation into RPE cells were discussed.
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BACKGROUND: Studies have shown that miRNA-148a can promote human bone marrow mesenchymal stem cells to differentiate into mature cardiomyocyte-like cells, but the effect of miRNA-148a on the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells has not been reported. OBJECTIVE: To investigate the effect of miRNA-148a on the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells. METHODS: Human induced pluripotent stem cells differentiating into cardiomyocyte-like cells were divided into three groups. Cells in the control group were not treated. Cells in the low expression group were treated with miRNA-148a for 28 days, and those in the high expression group were treated with mimics of miRNA-148a for 28 days. In addition, human induced pluripotent stem cells cultured for 28 days were taken as the blank control group. CCK-8 was used to detect cell proliferation activity. qRT-PCR was used to detect the expression of miRNA-148a. Immunofluorescence staining and western blot analysis were performed to detect the expression of MHC and cTnT protein. RESULTS AND CONCLUSION: The expression of intracellular miR-148a mRNA and cell proliferation activity in the low expression group were lower than those in the blank control and control groups, while those in the high expression group were significantly higher than those in the other three groups (P < 0.01). There were no positive expression of MHC and cTnT in the blank control group. There were positive expressions of MHC and cTnT in the control, low expression and high expression groups. The expression of MHC and cTnT protein in the low expression group was significantly lower than that in the control group, and that in the high expression group was significantly higher than that in the other three groups (P < 0.01). These results suggest that miRNA-148a can promote the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells.
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BACKGROUND: Chronic kidney disease is a severe threat to human health with no ideal treatment strategy. Mature mammalian kidneys have a fixed number of nephrons, and regeneration is difficult once they are damaged. For this reason, developing an efficient approach to achieve kidney regeneration is necessary. The technology of the combination of decellularized kidney scaffolds with stem cells has emerged as a new strategy; however, in previous studies, the differentiation of stem cells in decellularized scaffolds was insufficient for functional kidney regeneration, and many problems remain. METHODS: We used 0.5% sodium dodecyl sulfate (SDS) to produce rat kidney decellularized scaffolds, and induce adipose-derived stem cells (ADSCs) into intermediate mesoderm by adding Wnt agonist CHIR99021 and FGF9 in vitro. The characteristics of decellularized scaffolds and intermediate mesoderm induced from adipose–derived stem cells were identified. The scaffolds were recellularized with ADSCs and intermediate mesoderm cells through the renal artery and ureter. After cocultured for 10 days, cells adhesion and differentiation was evaluated. RESULTS: Intermediate mesoderm cells were successfully induced from ADSCs and identified by immunofluorescence and Western blotting assays (OSR1 + , PAX2 +). Immunofluorescence showed that intermediate mesoderm cells differentiated into tubular-like (E-CAD + , GATA3 +) and podocyte-like (WT1 +) cells with higher differentiation efficiency than ADSCs in the decellularized scaffolds. Comparatively, this phenomenon was not observed in induced intermediate mesoderm cells cultured in vitro. CONCLUSION: In this study, we demonstrated that intermediate mesoderm cells could be induced from ADSCs and that they could differentiate well after cocultured with decellularized scaffolds.
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Animals , Humans , Rats , Blotting, Western , Fluorescent Antibody Technique , In Vitro Techniques , Kidney , Mesoderm , Nephrons , Regeneration , Renal Artery , Renal Insufficiency, Chronic , Sodium Dodecyl Sulfate , Stem Cells , UreterABSTRACT
OBJECTIVE@#To investigate the molecular genetic mechanism of Charcot- Marie-Tooth (CMT) disease in a pedigree.@*METHODS@#Genomic DNA was extracted from the peripheral blood of the family members of a pedigree with autosomal dominant CMT disease, and 65 candidate genes of the proband were screened using target exon capture and the next generation sequencing, and the suspicious genes were verified using Sanger sequencing. PolyPhen-2, PROVEAN and SIFT software were used to predict the function of the mutant genes, and PyMOL-1 software was used to simulate the mutant protein structure.@*RESULTS@#A heterozygous missense mutation [c.371A>G (p.Y124C)] was detected in exon 3 of gene of the proband. This heterozygous mutation was also detected in both the proband's mother and her brother, but not in her father. Multiple sequence alignment analysis showed that tyrosine at codon 124 of GDAP1 protein was highly conserved. All the 3 prediction software predicted that the mutation was harmful. Molecular structure simulation showed a weakened interaction force between the amino acid residues at codon 124 and the surrounding amino acid residues to affect the overall stability of the protein.@*CONCLUSIONS@#The mutation of gene may be related to the pathogenesis of autosomal dominant AD-CMT in this pedigree. The newly discovered c.371A>G mutation (p.Y124C) expands the mutation spectrum of gene, but further study is needed to clarify the underlying pathogenesis.
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Female , Humans , Male , Amino Acids , Charcot-Marie-Tooth Disease , Genetics , Genes, Dominant , Genetics , Heterozygote , High-Throughput Nucleotide Sequencing , Methods , Mutation, Missense , Nerve Tissue Proteins , Genetics , Pedigree , SoftwareABSTRACT
Objective@#To evaluate the feasibility of differentiation of human adipose mesenchymal stem cells (hADSCs) into endothelial-like cells and the safety of induced cells intraocular application.@*Methods@#HADSCs were induced to endothelial-like cells in vitro.The experimental group was added with streptomycin, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF), and the control group was added with the same amount of phosphate buffered saline (PBS), the expression of von Willebrand factor (vWF), a marker of endothelial cells, was observed in the two groups.Six C57BL/6J mice were randomly divided into experimental group and control group by using the random number method, 3 mice for each group, the induced cells were injected intravitreally into C57BL/6J mice as experimental group, and the same amount of PBS buffer was injected intravitreally into C57BL/6J mice as control group.The changes of retinal cells were observed by electron microscopy.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.@*Results@#After induction of hADSCs in vitro, the endothelial cell marker was expressed.VWF immunofluorescence of the experimental group showed strong green fluorescence, and the color rendering rate was 100%.The control group showed no coloration of vWF immunofluorescence, and the color positive rate was 0.After 1 month of intravitreal injection, the retinal ganglion cells and rod cells were not degraded and necrotic.@*Conclusions@#HADSCs can differentiate into endothelial-like cells in vitro, and there is no retinal toxicity in a short-term.
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Objective: Inducing human amniotic membrane mesenchymal stem cells (hAMSCs) to Schwann cells-like cells (SCs-like cells) in vitro, and to evaluate the efficacy of transplantation of hAMSCs and SCs-like cells on nerves regeneration of the rat flaps.
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Objective To investigate the effects of mannan-binding lectin ( MBL) on the differen-tiation of Th17 cells (T helper cell 17, Th17). Methods CD4+T cells were separated in vitro from fresh human cord blood by MACS ( magnetic-activated cell sorting ) separator. Anti-CD3 monoclonal antibody ( McAb) and anti-CD28 McAb were used to stimulate CD4+T cells with IL-6 and TGF-β1 as inducers. Then, these cells were treated with ( MBL group) or without ( control group) different concentrations of MBL. Percentages of Th17 cells in different groups were detected by fluorescence-activated cell sorting( FACS) after 72 hours of culturing. Quantitative real-time PCR ( Q-PCR) was used to analyze the expres-sion of RORγt ( retinoid-related orphan receptor-γ-t) at mRNA level in both control and MBL groups. En-zyme-linked immunosorbent assay (ELISA) was performed to detect the levels of IL-17A in supernatants of cell culture from different groups. FACS was used to detect the percentages of Th17 cells in MBL-/-and wild type ( WT) mice. Results MBL could significantly reduce the percentage of Th17 cells after 72 hours of culturing as compared with the control group. Moreover, MBL could significantly down-regulate the expres-sion of RORγt at mRNA level and decrease the expression of IL-17A. Results of animal experiments showed that the percentages of CD4+RORγt+Th17 cells in MBL-/- mice were significantly higher than those in WT mice. Conclusion MBL can inhibit the differentiation of CD4+T cells to Th17 cells, which is induced by IL-6 and TGF-β1.
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Objective To investigate the effects of mannan-binding lectin ( MBL) on the differen-tiation of Th17 cells (T helper cell 17, Th17). Methods CD4+T cells were separated in vitro from fresh human cord blood by MACS ( magnetic-activated cell sorting ) separator. Anti-CD3 monoclonal antibody ( McAb) and anti-CD28 McAb were used to stimulate CD4+T cells with IL-6 and TGF-β1 as inducers. Then, these cells were treated with ( MBL group) or without ( control group) different concentrations of MBL. Percentages of Th17 cells in different groups were detected by fluorescence-activated cell sorting( FACS) after 72 hours of culturing. Quantitative real-time PCR ( Q-PCR) was used to analyze the expres-sion of RORγt ( retinoid-related orphan receptor-γ-t) at mRNA level in both control and MBL groups. En-zyme-linked immunosorbent assay (ELISA) was performed to detect the levels of IL-17A in supernatants of cell culture from different groups. FACS was used to detect the percentages of Th17 cells in MBL-/-and wild type ( WT) mice. Results MBL could significantly reduce the percentage of Th17 cells after 72 hours of culturing as compared with the control group. Moreover, MBL could significantly down-regulate the expres-sion of RORγt at mRNA level and decrease the expression of IL-17A. Results of animal experiments showed that the percentages of CD4+RORγt+Th17 cells in MBL-/- mice were significantly higher than those in WT mice. Conclusion MBL can inhibit the differentiation of CD4+T cells to Th17 cells, which is induced by IL-6 and TGF-β1.
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Objective To investigate the differences in eNOS gene expression,activity and its metabolites before and after human bone marrow mesenchymal stem cells (hBMSCs) are induced into vascular endothelial cells.Methods hBMSCs were induced into vascular endothelial cells.The morphological changes of the cells were observed under inverted microscope.Transwell assay was used to detect the cells' migration ability.The protein expression of eNOS was detected by immunofluorescence and Western blot.The activity of eNOS was detected by ELISA and the content of NO in cell culture supernatant was determined by nitrate reduction method.Results Compared with those in the undifferentiated group,the morphological changes of the differentiated cells were obvious.Cell migration ability increased by 238.10％ (73.000±7.002 vs.21.000±4.359,P＜0.05).The expression of eNOS protein increased by 114.72％ (0.423±0.011 vs.0.197±0.079,P＜0.05).The activity of eNOS was enhanced by 157.49％ (4.967±0.073 vs.1.929±±0.103,P＜0.05).The synthesis and release of NO increased by 155.67％ (184.909±1.853 vs.72.323±0.426,P＜0.05).Conclusion After hBMSCs are induced into endothelial cells,the expression of eNOS gene increases,their activities increase,synthesis and release of the metabolite NO increase.It may provide a basis for prevention and treatment of cardiovascular diseases with stem cells.
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Objective To report the clinical and peripheral neuropathological findings in two patients with autosomal recessive Charcot-Marie-Tooth disease 2K(AR-CMT2K).Methods Case one was a nine year-old girl.She had distal weakness of lower limbs for six years, with calf atrophy and contracture of Achilles tendon for three years.Case two was an eight year-old boy.He had distal weakness of lower limbs with contracture of Achilles tendon and calf muscle atrophy for three years, and proximal weakness of low limbs for two years.The motor nerve conduction velocities in median nerves were 48.1 m/s in case one and 47.6 m/s in case two.The compound motor action potential amplitude of median nerves decreased by 46% in case one and 69% in case two.Sural nerve biopsies and gene targeted next-generation sequencing were performed in both patients.Results Density of myelinated fibers was 8 407/mm2 in case one and 7 714/mm2 in case two.The ratio of myelinated fibers with diameter over 8 μm was 2.6% in case one and 0 in case two.Both patients had small regenerating cluster of myelinated fibers.Thin myelinated fibers appeared in case one.In case two, atypical onion bulb formations with focal folded myelin appeared, and electromicroscopy revealed mitochondrial aggregate in axons.Compound heterozygous mutations of ganglioside-induced differentiation associated protein 1 gene were detected in both patients, including c.767A>G(p.H256R) and c.466G>A (p.A156T) in case one and c.767A>G and 845G>A(p.R282H) in case two.Conclusions Contracture of Achilles tendon may appear in early childhood of AR-CMT2K patients.The main pathological changes in sural nerve are loss of large myelinated fibers, mitochondrial aggregate in axons and myelin abnormalities.
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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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Aim To explore the proteomics mechanism of the differentiation induction effect of 4-amino-2-trif-luoromethyl-phenyl retinate(ATPR)on human leukemi-a K562 cells. Methods Human leukemia K562 cells were incubated with the same concentration (1 × 10 - 6 mol·L - 1 ) of ATPR or ATRA for 48 hours. The total cell proteins were collected, purified and digested by trypsin, solid phase extraction, and the peptides were detected by ESI-LC-MS / MS. The difference of the pro-tein expression between the cells treated with ATPR and ATRA was compared by using the Discoverer Pro-teome 1. 2 software, and the molecular function, the biological process and other information of those pro-teins were analyzed based on the DAVID, KEGG, STRING databases. Results 120 specific proteins were identified only in the ATPR group, 143 only in the ATRA group, and 422 other proteins in both groups. Results of DAVID analysis showed that ATPR-induced specific proteins were mainly involved in 39 biological processes of proteins and macromolecules metabolism, protein transport and localization and so on. Results of KEGG analysis revealed that ATPR-in-duced proteins participated in signal pathways, mainly metabolic pathways, PI3K-Akt signal pathway, TGF-beta signal pathway and other pathways in cancer. String protein interaction network analysis displayed that ATPR-induced proteins, like EIF3A, EIF6, RPL3, RPL8, RPL13, RPL7A, RPL21, RPS3, RPS14, NACA, BTF3, NHP2L1, PPP2CA proteins had direct interactions with more than or equal to 10 associated proteins. Conclusion The differentiation induction effect of ATPR on K562 cells might be as-cribed to the ATPR-induced proteins interaction net-work and the specific central proteins it induced, which are involved in the regulation of cell prolifera-tion, differentiation and apoptosis.
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Objective To establish the method of isolation,culture and identify biological characterization of mesenchymal stem cells from human umbilical cord (hUCMSCs);and study their multiple differentiation potency.Methods Stem cells from human umbilical cord were cultured by enzyme Wharton jelly method in vitro.The surface markers were identified by flow cytometry.Multi-differentiation capacity was identified by osteogenic and adipogenic differentiation.ALP was detected with Calcium cobalt staining.The mineralized ability in vitro was measured with Alizarin red staining.Theadipocyte differen-tiation ability was measured with oil red-O staining.Results Flow cytometry analysis revealed that CD73 (92.45%),CD90 (95.45%)and CD105 (96.45%)were highly expressed on these cells’surface,while CD34 (1.07%)were negative ex-pressed.Cells were cultured with induced-osteogenic medium after 3 weeks,ALP staining in the cytoplasm of black parti-cles,and a large amount of mineralized nodules within cells was observed after 4 weeks.Cells were cultured with induced-adi-pogenic medium after 2 weeks,the majority of these cells were round,oil red O staining of lipid droplets generated within cells was observed.Conclusion Mesenchymal stem cells from human umbilical cord have the potential of multi-directional differentiation.These cells could be induced to differentiate into adipocytes and osteoblasts,which laid the foundation for clinical stem cell therapy research source of seed cells.
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[ ABSTRACT] AIM:In order to observe the myocardial differentiation capacity of the dedifferentiated fat ( DFAT) cells treated with vitamin C in vitro.METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture.The DFAT cells of passage 3 were used in the study.Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks.The cell morphology was ob-served under microscope.The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot.RESULTS:Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture.The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence.After treated with neonatal rat heart cell lysate, the DFAT cells be-came cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure.The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells.The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C.No spontaneous beating cell was observed.CONCLUSION:Vitamin C enhances the differentiation of DFAT cells into cardio-myocyte-like cells.
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BACKGROUND:Currently, bone marrow mesenchymal stem cel s can differentiate into nerve cel s via many approaches. Different methods for inducing bone marrow mesenchymal stem cel s differentiating into nerve cel s have different ratios. OBJECTIVE:To investigate the difference between chemical method and co-culture method to induce the differentiation of rat bone marrow mesenchymal stem cel s into nerve cel s. METHODS:Rat bone marrow mesenchymal stem cel s were isolated and purified using whole bone marrow culture method, and then randomly divided into two groups:chemical group,β-mercaptoethanol was added;co-culture group, co-cultured in a Transwel chamber. RESULTS AND CONCLUSION:Visible protrusions from induced cel s showed radiation growth at 1 week of induced culture, and neuron-specific enolase staining was positive at 2 weeks of culture. Star-like structure of nerve cel s was visible in the co-culture group within 4-5 days of culture, and then more protrusions formed. Meanwhile, the positive rate of neuron-specific enolase was (70.82±2.46)%. After 6-7 days of culture, neuron-like cel s formed and were interconnected in the chemical group;while, the positive rate of neuron-specific enolase was (52.37±1.83)%. These findings suggest that cel microenvironment plays a leading role in the differentiation of bone marrow mesenchymal stem cel s into nerve cel s, and chemical induction method is inferior to the co-culture method.
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10.3969/j.issn.2095-4344.2013.23.011