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BACKGROUND:Cel replacement therapy as an effective strategy for reconstruction of the central nervous system has very broad application prospects. OBJECTIVE:To investigate the effect of stereotactic transplantation of neural stem cels into the brain on the neuromotor function of craniocerebral trauma rats. METHODS:Twenty male Sprague-Dawley rats were equivalently randomized into study and control groups. Animal models of craniocerebral trauma were made using the improved free-fal method in the rats. Then, model rats in the study and control groups were given parenchymal transplantation of embryonic neural stem cels and the same volume of culture medium with no stem cels at 1 day after injury, respectively. Neuromotor function of rats was assessed based on the neurological severity scores. At 2 weeks after transplantation, brain tissues were taken for hematoxylin-eosin staining, anti-BrdU, glial fibrilary acidic protein, β-tubulin III and tyrosine hydroxylase immunohistochemistry staining. RESULTS AND CONCLUSION:The neurological severity scores in the study group were significantly lower than those in the control group at 1 and 2 weeks after injury (P< 0.05). In the study group, there were many BrdU-positive neural stem cels in the brain tissues, some of which were positive for glial fibrilary acidic protein, β-tubulin III and tyrosine hydroxylase; while in the control group, there was no BrdU-positive cel in the brain tissues. Experimental findings show that neural stem cels stereotacticaly transplanted into the brain can proliferate and differentiate in the brain lesion, and thereby notably improve the neuromotor function of rats with craniocerebral trauma.
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BACKGROUND:Isoflurane cannot only induce a wide range of large neuronal apoptosis, but also inhibit hippocampal neurogenesis in neonatal rats, thereby resulting in hippocampus-dependent learning and memory defects. OBJECTIVE:To investigate the isoflurane effect on proliferation and differentiation of the hippocampal neural stem cels. METHODS:Twenty-six Sprague-Dawley rats were randomly divided into air group and isoflurane group (n=13 per group). Rats in the isoflurane group were subjected to 2.5% isoflurane inhalation for 3 minutes folowed by 1.5% isoflurane inhalation for 4 hours. Rats in the air group only breathed in air. After the intervention, blood glucose and arterial blood gas changes were detected in the two groups. Additionaly, rats in the two groups were given intraperitoneal injection of 5-bromodeoxyuridine before and after intervention. At 24 hours after the last injection of 5-bromodeoxyuridine, brain tissues were taken to make frozen sections for immunofluorescence staining. RESULTS AND CONCLUSION:There were no significant difference in pH, PaO2, PaCO2, HCO3, BE and SaO2 levels between the two groups (P> 0.05). Compared with the air group, the number of BrdU+ cels was significantly less in the isoflurane group (P < 0.05), while the number of NeuroD+/BrdU+ cels was significantly higher in the isoflurane group (P < 0.05). The incidence of adverse reactions was 23% in the isoflurane group, which was significantly higher than that in the air group (7.7%;P < 0.05). These findings indicate that isoflurane can inhibit the proliferation of neural stem cels in the hippocampal dentate gyrus, and promote their differentiation into neurons.
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BACKGROUND:It wil provide a new insight into the future application of bone marrow mesenchymal stem cels in the treatment of spinal cord injury and tissue engineering by studying the effect of activation of Wnt signaling pathway in the neuronal differentiation of bone marrow mesenchymal stem cels. OBJECTIVE: To detect the expression of related genes by gene chip technology during the neuronal differentiation of bone marrow mesenchymal stem cels. METHODS:Human bone marrow mesenchymal stem cels were isolated and purified, and passage 5 cels were obtained. GatewayTM technology was used to build lentiviral vectors that was used to transfect Wnt-1 into human bone marrow mesenchymal stem cels. Control, non-transduction and transduction groups were set in this study. Human bone marrow mesenchymal stem cels were then induced to differentiate into neurons. Cel morphology was observed under inverted phase contrast microscope. Gene chip was used to detect the regulation changes and the differential expression of related genes in the Wnt signaling pathway. RESULTS AND CONCLUSION: Under the scanning electron microscope, the transfected cels were found to have the similar morphology of neuron-like cels. Analysis by the gene chip hybridization technique showed that 3 287 genes were up-regulated and 4 215 genes were down-regulated in the signal pathway. In the Wnt signaling pathway, genes related to the nervous system development and differentiation were up- or down-regulated. It is verified that the Wnt signal pathway is activated via Wnt-1 transduction, and the downstream genes appear to have genetic transcription so as to promote the neuronal differentiation of human bone marrow mesenchymal stem cels.
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BACKGROUND:Previous studies have found that the expression level of miR-25 in differentiated P19 cels is significantly lower than that in undifferentiated P19 cels. However, the effect of miR-25 on cardiomyogenesis and the relevant mechanism remain unclear. OBJECTIVE: To explore the effect and mechanism of miR-25 on the differentiation of P19 cels into cardiomyocytes. METHODS:P19 cels were cultured and differentiated into cardiomyocytesin vitro. The expression of miR-25 in differentiated and undifferentiated P19 cels was detected by real-time PCR. miR-25-overexpressing P19 cels were constructed by lipofection transfection, and were used to investigate the effect of miR-25 on the differentiation of P19 cels into cardiomyocytes. MicroRNA target analysis tools were used to explore potential targets of miR-25, and dual luciferase reporter assay was used to identify whether the 3’UTR of Pax3 mRNA was a binding target of miR-25. In addition, we transfected P19 cels with Pax3 shRNAs to silence the expression of Pax3, and investigated the effect of Pax3 on the differentiation of P19 cels into cardiomyocytes. RESULTS AND CONCLUSION: Expression level of miR-25 in differentiated P19 cels was obviously down-regulated compared with that in undifferentiated P19 cels. miR-25 overexpression promoted the differentiation of P19 cels into cardiomyocytes. By target prediction analysis, we confirmed that Pax3 was a potential target gene of miR-25. Luciferase assay further confirmed that miR-25 targeted Pax3 directly. Moreover, knockdown of Pax3 promoted the differentiation of P19 cels into cardiomyocytes. Taken together, miR-25 promotes the differentiation of P19 cels into cardiomyocytes by targeting Pax3. These findings offer new clues and theoretical basis for cardiomyogenesis and prevention and cure of congenital heart disease.
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BACKGROUND:Studies have shown that the number of osteoblasts is often decreased after osteoporosis, and osteoblast replacement therapy becomes a new target for the treatment of osteoporosis. OBJECTIVE:To observe the osteogenic differentiation of human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate. METHODS:Mesenchymal stem cels were isolated and purified from adult bone marrow using human lymphocyte separation medium. The expression of cel surface markers was detected by flow cytometry. Cel ultrastructure was observed by transmission electron microscope. Then, the bone marrow mesenchymal stem cels were cultured in osteogenic induction medium containing dexamethasone, vitamin C andβ-glycerophosphate, and RT-PCR was used to detect the bone morphogenetic protein-2 mRNA expression after osteogenic induction. RESULTS AND CONCLUSION:A large number of adherent cels were visible as fibrous growth at 2 weeks after culture and strongly expressed CD44, CD29, but did not express CD34, CD45. These cels could be induced to differentiate into osteoblasts, and express bone morphogenetic protein-2 mRNA. Alizarin red staining and alkaline phosphatase staining were positive for the cels. These findings suggest that human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate can differentiate into osteoblasts, and has a potential for the treatment of osteoporosis.
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BACKGROUND:Active components of Astragalus have an antioxidant effect, which is considered to result in the neuron-like differentiation of bone marrow mesenchymal stem cels. OBJECTIVE:To investigate the effect of astragalus polysaccharides combined with basic fibroblast growth factor to induce the differentiation of bone marrow mesenchymal stem cels into nerve cels. METHODS:After 24 hours of pretreatment with basic fibroblast growth factor, passage 3 human bone marrow mesenchymal stem cels were cultured with astragalus polysaccharides for 1-3 days (combined group). Blank control group and basic fibroblast growth factor group were set up. Expression of neuron-specific enolase and nestin was detected using western blot or immunocytochemical staining. RESULTS AND CONCLUSION:The expression of neuron-specific enolase was higher in the combined group than the basic fibroblast growth factor group (P < 0.05). Expression of nestin was found in both basic fibroblast growth factor group and combined group, but the gray value was higher in the combined group than the basic fibroblast growth factor group (P < 0.05). These findings indicate that astragalus polysaccharides combined with basic fibroblast growth factor is better to induce the neuronal differentiation of bone marrow mesenchymal stem cels.
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BACKGROUND:In recent years, neural stem cels are considered to be ideal for the treatment of spinal cord injury, but the proportion of its natural differentiation into neurons in the host body is relatively low, which severely restricts the therapeutic effect on spinal cord injury. OBJECTIVE:To investigate the effect of erythropoietin on the differentiation of neural stem cels in vitro. METHODS:Under sterile condition, neural stem cels from the hippocampus of neonatal Wistar rats were isolated, cultured and identified by immunofluorescencein vitro. The third generation of neural stem cels were randomly divided into 0.5, 5, 50 U/mL erythropoietin groups and control group (with no erythropoietin). RESULTS AND CONCLUSION:Compared with the control group, the differentiation rate of neural stem cels was significantly improved in the 0.5, 5, 50 U/mL erythropoietin groups (P 0.05). These findings indicate that erythropoietin can effectively induce the differentiation of neural stem cels into neurons in vitro, and moreover, it can significantly improve the differentiation rate of neural stem cels into neurons.
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BACKGROUND:The use of mesenchymal stem cels in the field of tissue engineering for osteoarticular injury repair is a very promising tool since these cels are readily expandable and able to differentiate into chondrocytes. Abundant evidence suggests that microRNAs play critical roles in chondrogenic differentiation of mesenchymal stem cels. OBJECTIVE:To observe the chondrogenic effect of human bone marrow mesenchymal stem cels transfected with lentiviral vectors bearing miR-221-3p/222-3p inhibition, thereby provding new strategies for cartilage injury. METHODS: miRNA microarray technology was applied to detect microRNAs expression profiles at three different stages of chondrogenic differentiation induction after transforming growth factor-β3 treatment and verified by real-time fluorescence quantitative PCR (RT-qPCR). Human bone marrow mesenchymal stem cels were infected with lentivirus bearing miR-221-3p/222-3p inhibition. After co-suppressing the expression of miR-221/222-3p, cel counting kit-8 was used to determine the cel proliferation, the differentiation of bone marrow mesenchymal stem cels towards chondrocytes was verified by type II colagen protein expression through immunohistochemistry and glycosaminoglycan accumulation was also elevated by sarranine O staining. RT-PCR was used to detect type II colagen and aggrecan mRNA expression at 21 days of chondrogenic induction. RESULTS AND CONCLUSION: The expression of miR-221-3p/222-3p was inhibited after Lv-miR221-3p/222-3p inhibition co-transfected into bone marrow mesenchymal stem cels. microRNA microarray and RT-qPCR results showed that the expression of miR-221-3p/222-3p was declined significantly at the anaphase of chondrogenic differentiation. The expression levels of chondrogenic markers, Aggrecan and type II colagen were significantly increased in the miR-221-3p/222-3p inhibition group and cel proliferation was also inhibited significantly compared with non-transduced cels or transduced with the empty lentiviral vector group. miR-221-3p/222-3p knockdown in bone marrow mesenchymal stem cels could inhibit proliferation but promote chondrogenic differentiation of bone marrow mesenchymal stem cels.
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BACKGROUND:Studies have indicated that melatonin can promote the differentiation of adipose-derived stem cels into neurons, and the effect of melatonin on the osteoblasts form adipose-derived stem cels is rarely reported. OBJECTIVE:To observe the osteogenic differentiation of adipose-derived stem cels and the effect of melatonin on the bio-viability of differentiated cels. METHODS:(1) Adipose-derived stem cels were isolated and purified from the inguinal fat of Kunming mice by type I colagenase digestion and differential adhesion method, respectively. Immunohistochemical staining of CD44 was used as a quality control. (2) Osteogenic induction medium was added to induce osteogenic differentiation of passage 2 adipose-derived stem cels. Alkaline phosphatase staining and von Kossa method were combined to evaluate differentiation condition. (3) Melatonin at variable concentrations was added to treat mature osteocytes originated from adipose-derived stem cels and MTT was applied to determine their viability at 24 and 48 hours after culture respectively to find out optimal condition of melatonin treatment. (4) Melatonin at the optimal concentration was used to treat differentiated cels and detect alkaline phosphatase activity after 3 days and 6 days respectively. RESULTS AND CONCLUSION: (1) After seeding for 48 hours, most cels were adherent, and after 4 days, the cels displayed multiple shapes and colonies of different sizes formed. After subculture, cel morphology homogenized as spindle shape. Cels positive for CD44 were brownish yelow, and localized mainly on the cel membrane. (2) Differentiated cels were positive for von Kossa staining and black sediments scattered in the extracelular matrix. Alkaline phosphatase expressed positively, and brown-black particles, appeared within cels. (3) Melatonin supplement improved the viability of differentiated cels; and 1, 10 and 100 μmol/L was observed as the optimal concentrations both at 24 and 48 hours. (4) The intracelular alkaline phosphatatse activity was increased with time in al the groups (P < 0.05). Compared with the blank group, the intracelular alkaline phosphatase activity in Melatonin groups (1, 10 and 100 μmol/L) had nochanges at 3 days, but significantly increased at 6 days (P < 0.05). These findings indicate that melatonin can enhance the proliferation of osteocytes differentiated from adipose-derived stem cels, and improve the activity of intracelular alkaline phosphatase.
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BACKGROUND:The use of neural stem cels provides a new approach for nervous system functional reconstruction and nerve regeneration. How to solve the induced differentiation of neural stem cels is stil a research hotspot. OBJECTIVE:To investigate the differentiation of rat neural stem cels into neurons and dopaminergic neurons under induction of glial cel-line derived neurotrophic factor gene. METHODS:PcDNA3-GDNF-GFP plasmids were constructed and transferredvia lipidosome into rat embryonic neural stem cels. Differentiation of neural stem cels after transfection was identified under a fluorescence microscope. Immunofluorescent staining was used to detect expression of β-tubulin III and tyrosine hydroxylase. RESULTS AND CONCLUSION:At 3 days after transfection, green fluorescence was observed in the transfected cels that were globular. At 7 days after transfection, the proportion of neural stem cels differentiating into neurons and dopaminergic neurons were significantly increased. These results indicate that the glial cel-line derived neurotrophic factor gene can induce the differentiation of neural stem cels into neurons and dopaminergic neurons.
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BACKGROUND:Neural stem cels have the potential to differentiate into neurons and glial cels to replace the injured brain cels, so as to achieve the purpose of repairing nerve injury. OBJECTIVE:To observe the neuronal differentiation ability of cel subsets with stem cel characteristics in the adult rat meningeal tissues. METHODS:Under anesthesia, the meningeal tissues were obtained from adult Sprague-Dawley rats to make cel suspension folowed by inoculation and subculture. Then, the Nestin immunofluorescence staining was performed. The third generation cels were culturedin vitro with complete culture medium containing trichostatin A. After 7 days of induction, western blot assay was used to detect the expression of NF-200 and BM88 proteins in neural cels. RESULTS AND CONCLUSION: At 24 hours of culture, some spherical cels were suspended and some cels adherent. In addition, some spherical cels scattered gradualy formed the clone spheres, and the growth rate decreased with the increasing volume. The positive expression of Nestin was detected by immunocytochemistry staining, and the cel nucleus was stained blue by Hoechst staining. BM88 and NF-200 proteins were al expressed at 7 days of neural induction. These findings indicate that the cel subsets with stem cel characteristics in the adult rat meningeal tissues can differentiate into neurons after in vitro induction.
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BACKGROUND:Our previous studies have shown that a soft substrate has a significant effect on morphology and cytoskeleton of rat bone marrow mesenchymal stem cel. OBJECTIVE:To explore the effect of polyacrylamide gels as soft substrates with different elastic moduli on the chondrogenic differentiation of human synovial-derived mesenchymal stem cels. METHODS:The synovium was harvested from patients with osteoarthritis under sterile conditions, and primary human synovial-derived mesenchymal stem cels were separated using limiting dilution assay. The flow cytometry and multi-directional differentiation experiments were used to identify the cel surface markers and function of the human synovial-derived mesenchymal stem cels, respectively. The polyacrylamide gels with the elastic modulus of 0.4, 6, 30 kPa, which were made using various amounts of acrylamide and bis-acrylamide, were used to culture human synovial-derived mesenchymal stem cels under induction with transforming growth factor-β1 for 7 and 14 days. RT-PCR was used to test the expression of chondrogenic genes, type II colagen gene and cartilage acidic protein 1. The 6-wel cel culture plates served as controls. RESULTS AND CONCLUSION: The human synovial-derived mesenchymal stem cels showed different cel morphology in the different elastic modulus of polyacrylamide gels. The expression of type II colagen gene and cartilage acidic protein 1 were affected by the different elastic modulus of polyacrylamide gels and culture time, and there was an interaction between these two factors. At 7 days of induction, the expression of cartilage acidic protein 1 gene on 6 kPa polyacrylamide gels was the highest (F=44.350,P=0.000); meanwhile, the expression of type II colagen gene on 0.4 kPa polyacrylamide gels was the highest (F=6.384,P=0.005). These findings indicate that polyacrylamide gels with lower elastic modulus are superior to routine culture plates to promote the chondrogenic differentiation of human synovial-derived mesenchymal stem cels.
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BACKGROUND:With respect to mesenchymal stem cels from other sources, synovial mesenchymal stem cels are rich in source, and moreover, the synovial tissue can regenerate quickly after partial hepatectomy and lead to fewer complications, in recent year, which have become a hot spot in stem cel research. OBJECTIVE:To observe the proliferation and directional differentiation of synovial mesenchymal stem cels from osteoarthritis patients. METHODS:Synovial mesenchymal stem cels were isolated and cultured. MTT assay was used to detect cel proliferation ability. Alkaline phosphatase activity was detected quantitatively at 7 days of osteogenic induction, and osteogensis-related gene expression was measured at 7, 14, 21 days of osteogenic induction. Alizarin red staining was performed at 21 days of induction. RESULTS AND CONCLUSION:(1) Passage 3 synovial mesenchymal stem cels proliferated faster, which were in latent period at 1, 2 days after inoculation, in logarithmic growth phase at 3-6 days, and then entered into the plateau phase at 7 days. (2) The activity of alkaline phosphatase was significantly higher in the induction group than the control group at 3, 7, 10 days after osteogenic induction (P < 0.05). The cels were positive for alizarin red staining at 21 days of osteogenic induction, and there were calcium deposits and calcium nodules in the extracelular matrix. (3) Bone-binding protein and Runx2 were visible at 7 days of osteogenic induction, and reached the peak at 21 days. These findings indicate that synovial mesenchymal stem cels from patients with advanced osteoarthritis have strong proliferation ability, which can differentiate into osteoblasts under in vitro induction.
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BACKGROUND:In recent years, stem cel therapy for early osteonecrosis of the femoral head has become an alternative method, but the quality of stem cels is a key to the therapeutic outcomes. OBJECTIVE:To evaluate the proliferative ability and directional differentiation ability of autologous bone marrow mesenchymal stem cels in a rat model of steroid-induced femoral head necrosis. METHODS:Twenty Sprague-Dawley rats were randomly divided into control and observation groups with ten in each group. An animal model of steroid-induced femoral head necrosis was built in the observation group, and then bone marrow mesenchymal stem cels from rats in both two groups were isolated and cultured. Cel counting kit-8 was used to detect proliferation of passage 3 cels. Bone marrow mesenchymal stem cels at passage 3 were selected in the two groups for osteogenic and adipogenic induction. Alkaline phosphatase staining and alizarin red staining were adopted at 7 and 14 days of osteogenic induction, and oil red O staining as performed at 21 days of adipogenic induction. RESULTS AND CONCLUSION:The absorbance values of bone marrow mesenchymal stem cels were lower in the observation group than the control group at 1, 3, 5 days of culture, but there was no significant difference between two groups (P > 0.05). Until the 7th day of culture, the absorbance value and alkaline phosphatase activity in the observation group were significantly lower than that in the control group (P < 0.05). Additionaly, there were fewer calcium nodules and lipid droplets in the observation group compared with the control group. These findings suggest that the proliferative ability and directional differentiation ability of autologous bone marrow mesenchymal stem cels from a rat model of steroid-induced femoral head necrosis are both decreased.
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BACKGROUND:Mesenchymal stem cels have the capacity of self-renewal and differentiation into certain lineage cels under appropriate conditions. But many mechanisms are unknown until now. OBJECTIVE:To clarify the role of miR-302 in the regulation of osteogenic and adipogenic differentiation of adipose-derived mesenchymal stem cels. METHODS:Chemicaly synthesized miR-302 specific mimics were transfected into adipose-derived mesenchymal stem cels as experimental group. miR-NC, miR-302b negative control mimics, was transfected into another cels as control group. By the experiments of alkaline phosphatase staining, alkaline phosphatase activity assay, alizarin red staining, oil red O staining and extraction test, the effect of miR-302 upregulation on the adipogenic and osteogenic differentiation of adipose-derived mesenchymal stem cels was analyzed and compared. Western blot assay was used to detect the expression of Runx2 and alkaline phosphatase after regulation of miR-302. RESULTS AND CONCLUSION: (1) Overexpression of miR-302 decreased the precipitate and activity of alkaline phosphatase significantly as compared with the control group (P < 0.05). (2) Overexpression of miR-302 inhibited the formation of mineral deposits and calcium nodules, and the number of calcium nodules in the experimental group was significantly lower than that in the control group (P < 0.05). (3) The number of cels positive for oil red O staining was significantly higher in the experimental group than the control group, which further showed the absorbance values of oil red O staining in the experimental group obtained in the extraction test were significantly increased (P < 0.05). (4) At 6 days of osteogenic induction, the expressions of Runx2 and alkaline phosphatase in the experimental group were decreased to different extents. These findings indicate that overexpression of miR-302 can suppress osteogenesis and accelerate adipocytes generation of adipose-derived mesenchymal stem cels. miR-302 plays a two-way regulatory role to balance the osteogenic and adipogenic differentiation of mesenchymal stem cels.
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BACKGROUND:Numerous studies have confirmed that neovascularization plays an important role in the growth, invasion and metastasis of tumors. OBJECTIVE:To investigate the features of CD133+ ovarian cancer stem-like cels differentiating into vascular endothelial cels. METHODS:CD133+ ovarian cancer stem-like cels were successfuly harvested from A2780 ovarian cancer cel lines using serum-free culture method, and incubatedin vitro onto 96-wel plates with or without Matrigel. Then, we observed the capacity of CD133+ ovarian cancer stem-like cels and human umbilical vein endothelial cels to form tube-like structures at different time points. Through xenograft experiments, the role of CD133+ ovarian cancer stem-like cels in the angiogenesis of ovarian cancer was observed using immunofluorescence staining. RESULTS AND CONCLUSION:CD133+ovarian cancer stem-like cels and human umbilical vein endothelial cels cultured with no Matrigel had no corresponding lumen formation, and could not express CD31. But those cultured with Matrigel had lumen formation and expressed CD31 significantly. After tumor formation, human-derived CD31 expression was observed in the tumors. These findings indicate that CD133+ ovarian cancer stem-like cels can differentiate into vascular endothelial cels, and be involved in tumor revascularization.
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BACKGROUND:The induced concentration for osteoblasts is often introduced as reference to induce odontoblast differentiation. However, there are no reports on other concentrations. OBJECTIVE: To observe the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein during low-dose β-glycerophosphate-induced differentiation of dental pulp stem cels into odontoblasts. METHODS:Human dental pulp stem cels were isolated and cultured, and then induced by different concentrations of inducing solution to differentiate into adipocytes and osteoblasts, which could verify the multi-directional differentiation ability of human dental pulp stem cels. Under 5 mmol/L β-glycerophosphate, dental pulp stem cels differentiated into odontoblasts. At 7, 14, 21, 28 days of culture, RNA samples were extracted from dental pulp stem cels in each group, and reverse-transcription PCR was used to detect the expression of dentin matrix protein-1, dentin sialoprotein and matrix extracelular phosphoglycoprotein. Mineralized nodules were detected by alizarin red S staining to show the successfuly osteogenesis induction. RESULTS AND CONCLUSION: Human dental pulp stem cels could be induced to adipocytes and osteoblasts. The results of reverse-transcription PCR showed that the dental pulp stem cels under 5 mmol/L β-glycerophosphate could increase the expression of dentin matrix protein-1 and dentin sialoprotein, but downregulate the expression of matrix extracelular phosphoglycoprotein at 7, 14, 21 days. At 28 days of culture, dental pulp stem cels were al successfuly mineralized detected by alizarin red S. There were some red mineralized nodules. These findings indicate that the 5 mmol/L β-glycerophosphate can induce the differentiation of dental pulp stem cels into odontoblasts successfuly, up-regulate the mRNA expression of dentin sialoprotein and dentin matrix protein-1, and meanwhile down-regulate the mRNA expression of matrix extracelular phosphoglycoprotein.
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BACKGROUND:Adipose-derived stem cels are pluripotent stem cels developed from the mesoderm, which can differentiate into different lineages induced by specific growth factors and under certain environmental conditions. OBJECTIVE: To describe the induced differentiation and identification of adipose-derived stem cels in detail. METHODS:A computer-based search of Wanfang and PubMed databases was performed for relevant articles published from 2005 to 2014 using the keywords of “adipose derived stem cels, induced, differentiation” in Chinese and English, respectively. According to inclusion and exclusion criteria, 37 articles were selected in result analysis. RESULTS AND CONCLUSION:Adipose-derived stem cels can differentiate into chondrocytes under the induction of ascorbic acid, insulin, dexamethasone, and transforming growth factor β. Recipe for adipogenic induction medium consists of 3-isobutyl-1-methylxanthine (IBMX), insulin, dexamethasone, indomethacin; and commonly used inducers for osteogenic differentiation include dexamethasone or vitamin D3, ascorbic acid,β-glycerophosphate. Basic fibroblast growth factor, epidermal growth factor and vitamin B27 may be combined to induce the differentiation of adipose-derived stem cels into neurons. 5-Azacytidine acts as a commonly used factor for inducing the differentiation of adipose-derived stem cels into cardiomyocytes. Combination of vascular endothelial growth factor and basic fibroblast growth factor can induce adipose-derived stem cels to differentiate into vascular endothelial cels. With the rapid development of molecular biology and cel biology, research on the differentiation of adipose-derived stem cels wil be more in-depth. Based on the current observation of adipose-derived stem cel differentiation, internal molecular mechanism as wel as genes and proteins for regulation of adipose-stem cel plasticity should be explored in depth.
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BACKGROUND:Bone morphogenetic protein 9 is proved to promote the osteogenic differentiation of various kinds of stem cel s, but whether it can induce the osteogenic differentiation of dental fol icle cel s in vitro is yet unclear. OBJECTIVE:To investigate whether bone morphogenetic protein 9 can induce the osteogenic differentiation of rat dental fol icle cel s in vitro. METHODS:Purified rat dental fol icle cel s at passage 3 were transfected with bone morphogenetic protein 9 adenovirus. Then, alkaline phosphatase activity, calcium deposition and expression of osteogenesis-related factors at mRNA and protein levels were detected in the dental fol icle cel s. RESULTS AND CONCLUSION:After transfection with bone morphogenetic protein 9, the dental fol icle cel s showed continuously enhanced alkaline phosphatase activities and obviously enhanced calcium deposition. Real-time PCR results demonstrated that the mRNA expressions of alkaline phosphatase, osteocalcin, bone sialoprotein, osteopontin and core binding factor were increased significantly. The western blot assay showed that the expression of osteopontin enhanced in the dental fol icle cel s after transfection with bone morphogenetic protein 9. In summary, bone morphogenetic protein 9 can induce the osteogenic differentiation of dental fol icle cel s.
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BACKGROUND:Ten-eleven translocation (TET) family proteins are recently discovered DNA dioxygenases that convert methylcytosine to hydroxymethyl cytosine, which is essential for regulating cel proliferation and differentiation, but the expression pattern of TET family proteins in human dental pulp cel s is stil unclear. OBJECTIVE:To investigate the expression pattern of TET family proteins during the differentiation of human dental pulp cel s. METHODS:Cel ular distribution and expression of TET family proteins were determined by immunofluorescence in human dental pulp cel s that were cultured and isolated using digestion method. The protein levels of TETs during cel passage (P1-P7) were detected with western blot assay, and their potential changes during odontogenic induction (7 and 14 days) were confirmed using real-time quantitative PCR and western blot analyses at mRNA and protein levels, respectively. RESULTS AND CONCLUSION:Al TETs were expressed in the nucleus and the cytoplasm of human dental pulp cel s During serial cel passage, TET1 protein expression was increased until the 6th passage, TET2 significantly increased at the 2nd and 3rd passages and then decreased (P0.05). Both mRNA and protein expression levels of al TETs were elevated during odontogenic induction (P<0.05). These results indicated that TETs may contribute to cel differentiation of human dental pulp cel s.