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BACKGROUND:Recently, the role of mesenchymal stem cels in aplastic anemia has been widely explored. However, its underlying mechanism remains unclearly. OBJECTIVE: To study the effect of umbilical cord blood and bone marrow mesenchymal stem cels on hematopoietic support and secretory function of T lymphocytes in patients with aplastic anemia. METHODS: Cord blood and bone marrow samples from 48 cases of aplastic anemia and 48 healthy lying-in women to isolate mesenchymal stem cels using flow cytometry. Mesenchymal stem cels from the cord blood and bone marrow were respectively co-cultured with cord blood mononuclear cels to count burst forming units-erythroid and colony forming units-granulocyte/macrophage. Mesenchymal stem cels were co-cultured with T lymphocytes from aplastic anemia patients undergoing phytohemagglutinin stimulation, and ELISA was used to detect interleukin-2, interleukin-4 and interferon-γ levels secreted from T lymphocytes. RESULTS AND CONCLUSION:The number of burst forming units-erythroid and colony forming units-granulocyte/macrophage significantly increased in normal bone marrow or umbilical cord blood mesenchymal stem cels co-cultured with cord blood mononuclear cels (P < 0.05), but reduced remarkably in umbilical cord blood mesenchymal stem cels from aplastic anemia patients co-cultured with cord blood mononuclear cels (P < 0.05). Levels of interleukin-2, interleukin-4 and interferon-γ from T lymphocytes were inhibited significantly after co-culture with normal bone marrow mesenchymal stem cels compared with phytohemagglutinin-induced T lymphocytes (P < 0.05). There was a similar inhibitory effect after co-culture with normal umbilical cord blood mesenchymal stem cels. There was a significantly reduction in the capacity of inhibiting interleukin-2, interleukin-4 and interferon-γ levels from T lymphocytes after co-culture with bone marrow mesenchymal stem cels from aplastic anemia patients (P < 0.05). Aplastic anemia patients show some functional defects in their bone marrow mesenchymal stem cels that have a weaker inhibitory role than normal bone marrow or umbilical cord blood mesenchymal stem cels in the hematopoietic support and secretory function of T lymphocytes. These findings indicate that mesenchymal stem cels from aplastic anemia patients can influence the pathological progress through weakening hematopoietic support and secretory function of T lymphocytes. Cite this article:Li GC, Song YP, Zhang YJ, Li G, Wang H, Xie J.Effects of mesenchymal stem cels on hematopoietic support and secretory function of T lymphocytes in patients with aplastic anemia. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):107-112.
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BACKGROUND:In recent years, in-depth studies that single Chinese herbs or extracts, compound traditional Chinese medicine and medicated serum are used to regulate the directional differentiation of bone marrow mesenchymal stem cels into myofibroblasts, chondrocytes, osteoblasts, myocardial cels and nerve cels, which have become a highlight in the tissue engineering research. OBJECTIVE:To review the latest progress in the directional differentiation of bone marrow mesenchymal stem cels induced by Chinese herbs or their extracts. METHODS:The first author searched the CNKI, Wanfang and PubMed databases using the keywords of “Chinese herb, directional differentiation, mesenchymal stem cels” in Chinese and English, respectively, to retrieve relevant articles published from January 2010 to January 2016. Repetitive articles or those with no originality were eliminated. Totaly 99 articles were searched initialy, and then 43 articles were included in result analysis. RESULTS AND CONCLUSION:As the strongest seed cels in the bone differentiation system, bone marrow mesenchymal stem cels have a wide range of directional differentiation potential, and highlight the important value in combination with Chinese herbs for clinical treatment of various refractory diseases, especialy for treatment of metabolic bone diseases, bone defects, nonunion and delayed union, which is not only conducive to in-depth, multi-angle studies on effects and mechanisms of Chinese herbs, but also to clinical treatment of various refractory diseases using bone marrow mesenchymal stem cels. Cite this article:Li N, Li YF, Xie XW, Song M, Xu SH, Li DP.Directional differentiation of bone marrow mesenchymal stem cel induced by traditional Chinese Medicine. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):135-139.
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BACKGROUND:The microRNAs are involved in regulation of stem cel proliferation, differentiation and aging. To study the effect of Let-7c, a member of Let-7, on the neural differentiation of bone marrow mesenchymal stem cels provides new ideas for stem cel therapy. OBJECTIVE: To investigate the role of Let-7c in the neural differentiation of bone marrow mesenchymal stem cels. METHODS: The lentiviral vectors of Let-7c-up and Let-7c-inhibition were constructed and transfected into rat bone marrow mesenchymal stem cels. Optimal multiplicity of infection was screened. The cels were divided into non-transfected group, negative control group (transfected with empty virus), transfected enhancement group (transfected with LV-rno-Let-7c-up), transfected inhibition group (transfected with LV-rno-Let-7c-5p-inhibition). Bone marrow mesenchymal stem cels were treated with fasudil as an inducer for triggering the cels to differentiate into neurons. The fluorescence expressed by transfected cels was observed under inverted fluorescence microscope. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The cel viability was determined by MTT method. RESULTS AND CONCLUSION:Under the inverted fluorescence microscope, the cels were successfuly transfected with LV-rno-Let-7c-up and LV-rno-Let-7c-5p-inhibition. Fasudil induced bone marrow mesenchymal stem cels to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in the transfected enhancement group were significantly higher than those in the negative control group (P < 0.05), while in the transfected inhibition group, they were lower than those in the negative control group (P < 0.05). These findings indicate that the differentiation percentage of bone marrow mesenchymal stem cels is increased by fasudil after transfection with LV-rno-Let-7c-up, and Let-7c may promote the differentiation of bone marrow mesenchymal stem cels into neurons.
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BACKGROUND:Numerous studies have demonstrated that bone marrow mesenchymal stem cels can be induced to differentiate into myocardial cels under certain conditions. Dimethyl sulfoxide is one of the commonly used inducers, and its mechanism is mainly by inhibiting the c-myc gene expression, thus reducing endogenous poly(adenosine diphosphate nucleotide) level. OBJECTIVE:To study the feasibility of dimethyl sulfoxide inducing the myocardial differentiation of bone marrow mesenchymal stem cels and its optimal concentration. METHODS:Bone marrow mesenchymal stem cels from Sprague-Dawley rats were isolated and culturedin vitro, and then induced by dimethyl sulfoxide to differentiate into myocardial cels. According to the concentrations of dimethyl sulfoxide, there were three groups: 0.6%, 0.8% and 1.0% group. Additionaly, a blank control group with no induction was set up. After 72 hours of induction, induction media were removed, and cels were then cultured in normal media for 4 weeks. RESULTS AND CONCLUSION: Morphology and immunocytochemistry detection results confirmed that dimethyl sulfoxide could induce the differentiation of bone marrow mesenchymal stem cels into myocardial celsin vitro, and differentiated cels expressed desmin, α-actin, cTnT, cTnI and P38MAPK. The optimal induced concentration of dimethyl sulfoxide was 1.0%. Immunofluorescence double staining and electron microscope results further confirmed that dimethyl sulfoxide could induce the myocardial differentiation of bone marrow mesenchymal stem cels. Cite this article:Sun LY. Dimethyl sulfoxide induces differentiation of bone marrow mesenchymal stem cels into myocardial cels. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):26-30.
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BACKGROUND:Stem cels are induced to differentiate into endothelial-like cels that can be used for the treatment of diabetic lower extremity vascular disease. However, it is unclear whether these endothelial-like cels can completely replace endothelial cels to improve vascular disease and what are the differences between endothelial-like cels and endothelial cels. OBJECTIVE:To explore the differences and similarities between endothelial-like cels and human umbilical vein endothelial cels in the aspects of morphology, function, and viability. METHODS:Umbilical cord mesenchymal stem cels and umbilical vein endothelial cels were isolated, cultured and identified using flow cytometry and immunohistochemical method. Isolated umbilical cord mesenchymal stem cels were induced in DMEM-LG/F12 containing 10 μg/L vascular endothelial growth factor, 10 μg/L basic fibroblast growth factor and 2% fetal bovine serum to differentiate into endothelial-like cels folowed by immunohistochemical identification. To compare endothelial-like cels with human umbilical vein endothelial cels, cel migration detection, active substance measurement and three-dimensional angiogenesis test were performed. RESULTS AND CONCLUSION:Isolated umbilical cord mesenchymal stem cels strongly expressed the surface markers of mesenchymal stem cels, and human umbilical vein endothelial cels strongly expressed CD31 and VWF. After induction, the umbilical cord mesenchymal stem cels were identified to highly express CD31 and VWF. Through cel migration, active substance and three-dimensional angiogenesis tests, endothelial-like cels were similar to endothelial cels in the function and activity, and superior to endothelial cels. Cite this article:Hao XJ, Hao HY, Zhu MJ, Yuan Z, Li WW, Chen J, Zhu LY. Endothelial-like cels versus human umbilical vein endothelial cels. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):83-88.
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BACKGROUND:Currently, there is a lack of efficient, non-invasive way to transplant stem cels to the target organ or tissue. Exploring a way to guide targeting transplantation of stem cels and to improve the efficiency of stem cel homing is now one of focuses in the field of stem cels research. OBJECTIVE: To establish a simple and feasible method to chemicaly modify the cel surface using biotin-streptavidin reaction system, and to evaluate the efficiency of this method to label bone marrow mesenchymal stem cels (BMSCs) and its effects on cel biological functions. METHODS: Passage 3 BMSCs were obtained by whole bone marrow culture method and verified by flow cytometry. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were used to equip the adhesion molecule ligand, sialyated LewisX (SLeX), to the BMSCs surface. The labeling rate of BMSCs was assessed using fluorescence microscope, the vitality of BMSCs was evaluated by trypan blue staining, and the proliferation of BMSCs was evaluated by cel counting kit-8 assay. Adipogenic and osteogenic inductions were used to evaluate the effect of the method on the multi-differentiation function of BMSCs. RESULTS AND CONCLUSION: After culture for 2 weeks, passage 3 BMSCs were obtained and confirmed by expressing CD90, CD29 and lack of CD34, CD45. Biotin, streptavidin, sulfonated biotin-N-hydroxy-succinimide were successfuly used to equip sialyated LewisX (SLeX) to the BMSCs surface and had minor effects on the vitality, proliferation, and differentiation of BMSCs. This method was simple for surface modification and had a high modification rate of 88%. The homing of BMSCs modified by this method to the target organ or tissue could be greatly enhanced. Therefore, this method potentialy could have extensive and important applications.
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Abstract BACKGROUND: Alveolar bone deficiency wil not meet aesthetic and functional requirements for dental implants. OBJECTIVE:To observe the repair effect of passage 3 autologous bone marrow mesenchymal stem cels (BMSCs) and platelet-rich fibrin (PRF) on alveolar bone defects in rabbits. METHODS:Twenty-seven New Zealand rabbits were randomly divided into BMSCs/PRF group, PRF group and model group (n=9 per group). The left mandible incisors were extracted in al the rabbits under general anesthesia. BMSCs/PRF group was immediately implanted BMSCs/PRF composite into the alveolar socket, PRF group only implanted PRF, and model group implanted nothing. RESULTS AND CONCLUSION: In the model group, the alveolar crest and alveolar mucosa become sunken notably and narrowed. In the BMSCs/PRF and PRF groups, the thickness of alveolar bone wal, alveolar bone width, alveolar bone height difference, and bone mineral density were al increased, especialy in the former group. In addition, the trabecular arrangement was better in the BMSCs/PRF groups than the model and PRF group. Our findings indicate that alveolar socket filing with composite of BMSCs and PRF can achieve preservation of alveolar bone width and height after tooth extraction in rabbits.
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BACKGROUND:Ankylosing spondylitis is an autoimmune disease at high inflammatory state, and its pathogenesis is stil unclear. Besides, there is a lack of entirely satisfactory curative strategies. OBJECTIVE: To explore the immunoregulation capability of bone marrow mesenchymal stem cels from ankylosing spondylitis patients on macrophages and the potential therapeutic use of bone marrow mesenchymal stem cels from healthy donors on ankylosing spondylitis. METHODS: Bone marrow mesenchymal stem cels were extracted from 21 healthy donors and 25 ankylosing spondylitis patients respectively, and passage 4 cels were used in subsequent experiments. A human monocytic cel line was induced to differentiate into macrophages. The phenotypic markers of bone marrow mesenchymal stem cels and macrophages were detected by flow cytometry. Expressions of tumor necrosis factor-α and tumor necrosis factor-α-stimulated gene 6 (TSG-6) proteins in the supernatant of co-culture system were detected by ELISA. Quantitative real-time PCR was applied to detect the mRNA level of cytokines secreted by bone marrow mesenchymal stem cels and macrophages. RESULTS AND CONCLUSION:The typical mesenchymal stem cel surface markers were expressed in both bone marrow mesenchymal stem cels from healthy donors and patients with ankylosing spondylitis, and CD68 was detected positively in induced macrophages. The protein and mRNA levels of tumor necrosis factor-α secreted by macrophages co-cultured with bone marrow mesenchymal stem cels from patients with ankylosing spondylitis were obviously higher than those from healthy donors (P < 0.05). TSG-6 secreted by bone marrow mesenchymal stem cels from patients with ankylosing spondylitis was lower than that by bone marrow mesenchymal stem cels from healthy donors in both RNA transcriptional and protein levels (P < 0.05). Our study demonstrates that bone marrow mesenchymal stem cels from patients with ankylosing spondylitis shows abnormal immunoregulatory function on inhibiting the tumor necrosis factor-α secretion from macrophages, which reveals a mechanism of immune disorder in ankylosing spondylitis. The therapeutic mechanism of bone marrow mesenchymal stem cels from healthy donors may work by secreting enough TSG-6 to inhibit the activation of macrophages in patients with ankylosing spondylitis, and thereby to decrease the secretion of tumor necrosis factor-α. Cite this article:Sun SH, Wang P, Su CY, Xie ZY, Li YX, Li D, Wang S, Su HJ, Wu XH, Deng W, Wu YF, Shen HY. Bone marrow mesenchymal stem cels derived from patients with ankylosing spondylitis show abnormal immunoregulation capability on macrophages. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(1):13-19.
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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:We attempt to explore a low-cost, simple and effective way to cryopreserve bone marrow mesenchymal stem cels at-80℃. OBJECTIVE:To screen the optimal cryopreservation fluid for bone marrow mesenchymal stem cels and to verify the biological features of bone marrow mesenchymal stem cels after long-term cryopreservation. METHODS: Bone marrow mesenchymal stem cels were cultured using adherent method and the biological features and purity of cels were detected using immunofluorescence method. Bone marrow mesenchymal stem cels were cryopreserved in the cryoprotectant medium containing low-sugar DMEM, fetal bovine serum and dimethyl sulfoxide at different proportions at-80℃ for a short term. Then, the optimal cryoprotectant was selected to storage the bone marrow mesenchymal stem cels. After 1, 3, 6 months of cryopreservation, the cels were resuscitated, cultured and passaged. Passage cels were identified immunofluorescence method to determine the biological features of bone marrow mesenchymal stem cels cryopreserved at-80℃. RESULTS AND CONCLUSION:Cryoprotectant medium of 80% DMEM+10% fetal bovine serum+10% dimethyl sulfoxide was suitable for cryopreserving MSCs at -80℃, and resuscitated cels were able to proliferate in vitro, and passage normaly, indicating the cryopreserved bone marrow mesenchymal stem cels stil maintain the original biological activity.
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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:Infrapatelar fat pad is often partialy resected in the knee surgery, which can be used as an important source of adipose-derived mesenchymal stem cels. OBJECTIVE: To explore the strategies of isolation, culture, and identification of adipose-derived mesenchymal stem cels from the infrapatelar fat pad and to detect the expression of cel surface markers of human adipose-derived stem cels. METHODS: Infrapatelar fat pad was obtained from patients undergoing knee arthroscopy surgery, and attached cels were obtained from adipose tissue by using colagenase I. Cels were cultured in 10% low-sugar DMEM. Stem cels proliferation was detected by means of MTT and then, cel growth curve was made. The obtained cels were induced and differentiated into adipocytes and osteocytes. Expressions of cel surface markers CD29 and CD44 were detected. RESULTS AND CONCLUSION:A few of attached cels were observed after cultured 24 hours. Cels proliferated faster and exhibited spindle shape after 1 week. Cel adherence and proliferation were speeded up after subculture. Growth curve of cels exhibited that the passages 5 and 2 cels had higher reproductive activity than passage 8 cels. The obtained cels can be induced and differentiated into adipocytes and osteocytes. Results from flow cytometry showed that 96.8% passage 5 cels expressed CD29 and 97.6% expressed CD44. These findings indicate that high-purity adipose-derived mesenchymal stem cels with high reproductive ability are easy to be isolated from the infrapatelar fat pad, which may be a kind of ideal seed cels for cartilage tissue engineering.
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BACKGROUND:Since the FDA was the first to approve autologous bone marrow stem cel transplantation for treatment of myocardial infarction in 2003, there has a large number of clinical and basic research reports. However, their conclusions are different and stem cel homing is a key point. OBJECTIVE:To explore the homing abilities of different subgroups of mouse bone marrow mesenchymal stem cels in myocardial regeneration. METHODS:After mouse bone marrow mesenchymal stem cels were detected using a mouse cardiac stem cel surface differentiation antigen, four cel subgroups were separated on the basis of CD45 and CD31. The homing abilities of the four subgroups were assayed in a Transwel chamberin vitro. The different cel subgroups were injected into the model mice suffering from myocardial infarction for 48 hours. The mice were sacrificed at 48 hours, 96 hours, and 7 days after injection; the hearts were taken and analyzed through whole-body imaging and fluorescence intensity detection. RESULTS AND CONCLUSION:The SCA-1+/CD45+/CD31+ subgroup exhibited the strongest homing ability. The whole-body imaging indicated that the fluorescence intensity of SCA-1+/CD45+/CD31+ subgroup was higher than that of the other subgroups at 48 hours, 96 hours and 7 days after stem cel injection. The migration rate of SCA-1+/CD45+/CD31+ subgroup was also the highest. These findings indicate that the homing ability of the SCA-1+/CD45+/CD31+ subgroup of mouse bone marrow mesenchymal stem cels exhibit a homing trend to the damaged myocardial tissue.
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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:The self-renew and regeneration capacity of the injured spinal cord is thought to be limited. Accordingly, cel transplantation is one potential strategy for promoting functional recovery after spinal cord injury. OBJECTIVE:To explore the effects ofPTEN silencing on the biological properties of bone marrow mesenchymal stem cels, hoping to offer better seed cels for tissue engineering. METHODS:Bone marrow mesenchymal stem cels were transfected with specific siRNA-silencedPTEN gene using the liposome method, and then RT-PCR was used to detect the mRNA expression ofPTEN. Variation of biological properties ofPTEN-transfected cels were detected by the way of MTT assay, cel cycle analysis, and Transwel assay. RESULTS AND CONCLUSION:PTEN is expressed highly in bone marrow mesenchymal stem cels, which is successfuly interfered by siRNA.PTEN-silenced cels have stronger survival, proliferation and migration abilities, which become a kind of better seed cels for tissue engineering.
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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:Mesenchymal stem cels can secrete a variety of cytokines and growth factors that promote the survival of surrounding cels and play a paracrine role. OBJECTIVE:To investigate the effect of human umbilical cord mesenchymal stem cels on the proliferation and apoptosis of ectopic endometrial cels. METHODS:After isolation and culture, human umbilical cord mesenchymal stem cels and ectopic endometrial cels were co-cultured as observation group, and ectopic endometrial cels cultured alone served as control group. At 24, 48, 72 hours of culture, the proliferation and apoptosis of ectopic endometrial cels were detected by MTT and flow cytometry, respectively; RT-PCR was used to measure the expression ofPTEN gene in ectopic endometrial cels. RESULTS AND CONCLUSION:At 24, 48 and 72 hours, the proliferation of ectopic endometrial cels in the observation was inhibited significantly as compared with the control group, and the hypodiploid peak ratio also increased significantly (P < 0.05). Over time, the cel inhibition rate was gradualy declined, and there were significant differences at different time points (alP < 0.05). Compared with the control group, the expression of PTEN gene was up-regulated significantly in the observation group (P < 0.05). In the observation group, the expression ofPTEN gene at 48 and 72 hours was significantly higher than that at 24 hours (P < 0.05). These findings indicate that in the human umbilical cord mesenchymal stem cels can inhibit the proliferation of ectopic endometrial celsin vitro and promote their apoptosis by up-regulation ofPTEN mRNA expression.
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BACKGROUND:Cirrhosis is a long-term consequence of chronic hepatic injury, which has no effective therapy. Mesenchymal stem cels have been shown to play a potential role in the treatment of liver fibrosis/cirrhosis. OBJECTIVE:To investigate the therapeutic effect and mechanism of human umbilical cord-derived mesenchymal stem cels on CCl4 induced liver fibrosis/cirrhosis in rats. METHODS:A CCl4-induced liver fibrotic/cirrhotic rat model was used, and human umbilical cord-derived mesenchymal stem cels were injectedvia the tail vein after modeling. Liver biochemical profile was measured by Beckman Coulter analyzer. Histopathological changes were assessed by Sirius red staining. The expressions of colagen type I, colagen type III, matrix metaloproteinases-2 and tissue inhibitor of matrix metaloproteinases-2 protein and mRNA in liver tissues were observed by immunohistochemistry, western blot and real-time PCR, respectively. RESULTS AND CONCLUSION:Liver biochemical profile indicated the transplantation of human umbilical cord-derived mesenchymal stem cels could improve the liver function of rats with liver fibrosis and cirrhosis. After cel transplantation, except 1-week cel transplantation group, the expressions of the matrix metaloproteinases-2 mRNA and protein were significantly increased, while the expressions of colagen type I, colagen type III and tissue inhibitor of matrix metaloproteinases-2 mRNA and protein significantly decreased, compared with the corresponding model groups. Human umbilical cord-derived mesenchymal stem cels play a role in the treatment of liver fibrosis and cirrhosis through upregulating the expression of matrix metaloproteinases-2 and lowering the expression of inhibitor of matrix metaloproteinases-2. With the continued presence of pathogenic factors, human umbilical cord-derived mesenchymal stem cel transplantation cannot reverse liver fibrosis or cirrhosis, and only delay the process of liver fibrosis or cirrhosis.
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BACKGROUND:Xenogeneic ligament is readily available, which has ligament scaffold structure and is conducive to tissue ingrowth and creeping substitution. After processing, the xenogeneic ligament, with the presence of good growth scaffold function, can be completely eliminated antigenicity that can cause immune rejection. OBJECTIVE:To study the feasibility of biological xenogeneic ligament graft instead of alogeneic ligament graft for reconstruction of goat anterior cruciate ligament. METHODS: Twenty-four healthy adult goats were randomly divided into A, B, C groups, and then, the left knee joints of goats were removed to establish animal models of anterior cruciate ligament injury in the three groups. After the establishment of the tibia and femoral bone tunnel, groups A, B, C were respectively transplanted with biological xenogeneic ligament graft combined with bone marrow mesenchymal stem cels, biological xenogeneic ligament graft alone, and alogenic ligament. RESULTS AND CONCLUSION: After implantation, no rejection but good biocompatibility was found in the group A, in which, the transplanted ligament as a functional scaffold for anterior cruciate ligament reconstruction, and new tissues in-grew and replaced the scaffold under intraarticular environment to form the new ligament with good bone tendon healing. However, there were no differences in histology, immune response, biomechanical findings between groups A and C. Additionaly, in the group A, the host tissues were found to grow into the scaffold and establish a micro-circulation. These findings indicate that xenogeneic ligament combined with bone marrow mesenchymal stem cels can accelerate the establishment of micro-circulation and promote the growth of ligaments, especialy improve the ligament revascularization significantly, but has no influence on the biological characteristics of the ligament.
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BACKGROUND:Induced pluripotent stem cels and mesenchymal stem cels-derived microvesicles have been confirmed in various tissue repairs, which are expected to become more effective and safe therapy for articular cartilage repair. OBJECTIVE:To overal understand the research progress in the use of induced pluripotent stem cels and mesenchymal stem cels-derived microvesicles in articular cartilage repair. METHODS: A computer-based search of PubMed and CNKI was performed by the first author for articles related to stem cel treatment of osteoarthritis published from 2003 to 2015. The keywords were “articular cartilage injury, bone marrow mesenchymal stem cels” in English and Chinese, respectively. In the same field, articles published recently or in authorized journals were preferred. RESULTS AND CONCLUSION:Articular cartilage injury is stil a difficulty in the orthopedics. Many repair methods have been reported, but they al have limitations. Induced pluripotent stem cels and mesenchymal stem cels-derived microvesicles bring a new hope for patients with articular cartilage injury. However, there are stil many problems to be solved, such as extracting and purifying a large amount of cels, proliferation and differentiation potentials, and mechanism underlying cartilage repair.