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1.
BACKGROUND: Choosing an effective means to label and trace the distribution, differentiation and migration of cells in vivo help to further explore the specific mechanism of cells that exert a therapeutic effect.OBJECTIVE: To understand the migration and localization of BrdU-labeled human umbilical cord mesenchymal stem cells in brain injury model rats.METHODS: Human umbilical cord blood samples were obtained, and the isolation of human umbilical cord mesenchymal stem cells was carried out. The primary and passage culture were performed. The phenotype of cells was detected by flow cytometry. Passage 3 human umbilical cord mesenchymal stem cells were labeled using BrdU, and the cell proliferation was detected using MTT method. BrdU-labeled cells were injected into brain injury rats via the tail vein. At 14 days after transplantation, brain tissues in the injury region were cut into sections and the migration and location of the umbilical cord mesenchymal stem cells were observed under inverted fluorescence microscope.RESULTS AND CONCLUSION:Cell surface specific markers CD45 and CD34 were detected by flow cytometry, but the cells could not express CD44, CD105 and CD29. Based on the cell growth curve, the cells came into a conditioning period at 1-3 days of seeding and came into a logarithmic phase at 3-5 days. BrdU-positive cells were visible at the injury region after 14 days, indicating that in the rats, transplanted human umbilical cord mesenchymal stem cells migrated from the peripheral blood to the site of brain injury to achieve the effective repair of injured parts.
中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程 相似文献
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BACKGROUND: Bone marrow mesenchymal stem cells have been widely used in the clinical treatment of neurodegenerative-related diseases, but their efficacy is reduced by low cell survival and migration rates at the site of injury. OBJECTIVE: To investigate whether miR-31 can enhance the migration and proliferation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells from C57BL/6 mice were cultured and identified, and the cells were divided into control, miR-31 agomir, and miR-31 antagomir groups. Bone marrow mesenchymal stem cells at passage 3 were inoculated in six-well plates (1×105/well). When the cells reached 50%-80% fusion, miR-31 agomir and miR-31 antagomir were added to the six-well plates after dilution with serum-free DMEM/F12. After 24 hours of transfection, the proliferation level of cells was analyzed by CCK-8 assay as well as the migration ability of cells was analyzed by Transwell assay. Protein expression of matrix metalloproteinase 2 and CXC chemokine receptor 4 was detected by western blot assay. RESULTS AND CONCLUSION: (1) miR-31 was successfully transfected with bone marrow mesenchymal stem cells without transfection reagents and emitted red fluorescence. (2) After transfection, the proliferation ability of cells in the miR-31 agomir group was enhanced compared with the control group, which increased proportionally with time (P < 0.05). Compared with the control group, miR-31 promoted the migratory ability of bone marrow mesenchymal stem cells in the miR-31 agomir group (P < 0.05) and also upregulated protein expression of matrix metalloproteinase 2 and CXC chemokine receptor 4 (P < 0.05). (3) The results indicated that miR-31 could improve the proliferation ability of bone marrow mesenchymal stem cells and promote the migration of bone marrow mesenchymal stem cells, which provides a basic study for efficient targeting of mesenchymal stem cell migration to the site of injury. © 2023, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved. 相似文献
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BACKGROUND: Immunotherapy with autologous immune cells has been developed as a major adjuvant therapy for malignant tumors, but its mechanism of action has not been elucidated.
OBJECTIVE: To investigate the relationship between cytokine-induced killer cell secretion and apoptosis in human liver cancer stem cells.
METHODS: Human liver cancer stem cells, HepG2 cells, were isolated and enriched using serum-free suspension method. The peripheral blood mononuclear cells from patients with liver cancer were induced by γ-interferon, CD3 monoclonal antibody and recombinant human interleukin-2 to form killer cells. Passage 1 liver cancer stem cells were divided into control group (culture alone) and experimental group (co-culture of cytokines-induced killer cells and human liver cancer stem cells). At 48 hours after culture, apoptosis in human liver cancer stem cells was detected using flow cytometry, and expression of caspase-3 mRNA and protein was detected using RT-PCR and western blot, respectively.
RESULTS AND CONCLUSION: The apoptotic rate in the control group was significantly lower than that in the experimental group (P < 0.05). The expressions of caspase-3 at mRNA and protein levels were both higher in the experimental group than the control group (P < 0.05). Experimental findings show that cytokines-induced killer cells can significantly promote apoptosis in human liver cancer stem cells, and up-regulate the caspase-3 mRNA and protein expressions dramatically.
中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程 相似文献
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BACKGROUND: Bone marrow mesenchymal stem cells have a low survival rate after implanted into the ischemic myocardium. However, hypoxia preconditioning (HPC) may enhance bone marrow mesenchymal stem cell proliferation and promote its survival rate.
OBJECTIVE: To explore whether Pim-1 is involved in HPC protecting against apoptosis of bone marrow mesenchymal stem cells and the relevant mechanism.
METHODS: Bone marrow mesenchymal stem cells were respectively subjected to HPC for 0, 6, 12, and 24 hours. The expression of Pim-1 and apoptosis-related genes were detected by RT-qPCR and western blot. Then, the best hypoxic preconditioning time was determined as 12 hours. Then, bone marrow mesenchymal stem cells were assigned to one of the following groups: control (without HPC), 12-hour HPC, 12-hour HPC+Pim-1 inhibitor groups. Flow cytometry analysis was used to detect the cell apoptosis, Transwell assay to analyze the cell migration ability in each group, and JC-1 kit to detect mitochondrial membrane potential. Animal models of myocardial infarction were established. One week after modeling, bone marrow mesenchymal stem cells were given via multi-point injection around the infarct zone of rats. Two weeks after modeling, heart tissues of rats were taken and sliced followed by DiI staining to calculate the survival rate of bone marrow mesenchymal stem cells. Additionally, rat cardiac function was assessed by echocardiography prior to and after modeling as well as at 4 weeks after cell transplantation.
RESULTS AND CONCLUSION: At 12 hours after HPC, the expression of Pim-1, p-Akt and Bcl-2 gene in the infarct region was significantly increased, but the expression of caspase-3 and Bax was significantly decreased. Compared with the control group, cell viability in the 12-hour HPC group was increased very significantly at 1 week after cell transplantation (P < 0.001), the migration and anti-apoptosis ability were enhanced significantly (P < 0.01) and the cardiac function of rats was significantly improved in the 12-hour HPC group (P < 0.05). All of these protective effects were blocked by the Pim-1 inhibitor. These findings indicate that HPC can protect bone marrow mesenchymal stem cells from apoptosis through activating Akt and up-regulating Pim-1, and thereby improve the therapeutic effect of bone marrow mesenchymal stem cell transplantation on ischemic heart diseases.
中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程 相似文献
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BACKGROUND: Thinking from ovarian cancer stem cell theory shows that: in the tumor cells, there are a fraction of stem cells with self-renewing ability and multipotent differentiation, which are the root causes of ovarian cancer recurrence and drug resistance. Studies have shown that CD90 can be used as a surface marker of mesenchymal stem cells and stem cells of other cancers.OBJECTIVE: To explore the biological features of CD90+ tumor cells from ovarian cancer tissues.METHODS:Primary ovarian cancer cells were isolated from the abdominal dropsy of ovarian cancer patients to sort CD90+ and CD90- cells using flow cytometry. RT-PCR was used to detect expressions of stem cell-related genes and epithelial to mesenchymal transition-related genes. Cell invasion was observed by Transwell invasion assay, cell proliferation and differentiation observed by clone formation assay, stem cell potential observed by suspension sphere-forming assay, and tumor formation rate observed by in vivo tumorigenicity experiment.RESULTS AND CONCLUSION: Compared with the CD90- cells, the expressions of CD44, CD133, ALDH1, N-cad and Vimentine were significantly higher in the CD90+ cells (P < 0.05), but the expression of E-cad was significantly decreased in the CD90+ cells (P < 0.05). Tumor formation rates of CD90- and CD90+ cells were increased significantly with the increase of seeded cell number, which was more obvious in CD90+ cells. The number of transmembrane cells, the number of cell clones and the number of suspended spheres were significantly higher in the CD90+ cells than the CD90- cells (P < 0.05). Experimental findings from this study show that CD90+ cells highly express epithelial to mesenchymal transition-related genes and stem cell-related genes, with higher invasion, proliferation and differentiation, in vivo tumorigenicity and potential of stem cells. CD90+ cell separation may be a new method to separate ovarian cancer stem cells.
中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程 相似文献
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BACKGROUND: Currently, conventional treatment methods for heart failure are all ineffective.
OBJECTIVE: To explore the combined effects of human umbilical cord stem cells and CD133+ cells in mice with heart failure, providing a new insight into the treatment of heart failure.
METHODS: Full-term newborn umbilical cord from vaginal delivery was collected to isolate CD133+ cells and human umbilical cord stem cells using lymphocyte separation medium method. Twenty Balb/C nude mice were randomly subjected to mononuclear cell injection (mononuclear cell group) or injection of CD133+ cells combined with human umbilical cord stem cells (combined group) via the tail vein after establishing heart failure models.
RESULTS AND CONCLUSION: Fourteen days after injection, the body weight and liver, heart and lung mass of mice were significantly larger in the combined group than the mononuclear cell group (P < 0.05). After 30 days, myocardial cells arranged regularly in the combined group, but disorderly in the mononuclear cell group; compared with the mononuclear cell group, the average area of myocardial collagen fibers was significantly decreased in the combined group (P < 0.05), and the level of serum matrix metalloproteinase-9 was also significantly lower in the combined group (P < 0.05). Masson staining showed that blue-stained collagen fibers in the combined group were less but arranged neatly; however, in the mononuclear cell group, the number of collagen fibers that arranged irregularly was increased to different extents. To conclude, the combined use of CD133+ cells and human umbilical cord stem cells has desired outcomes in the treatment of heart failure in mice, indicating a higher clinical value.
中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程 相似文献
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BACKGROUND: The periodontal ligament stem cells can promote periodontal tissue regeneration, providing a new way for the treatment of periodontitis.OBJECTIVE: To observe the inflammatory microenvironment effects on the biological properties of periodontal ligament stem cells.METHODS:Periodontal ligament stem cells from healthy controls and patients with periodontitis were primarily cultured by tissue digestion method, purified using limited dilution method, and identified through detection of CD146 and STRO-1. Then, passage 3 cells were taken and denoted as normal control and inflammation groups followed by osteogenic induction.RESULTS AND CONCLUSION: Purified cells from two sources both expressed STRO-1 and CD146. Periodontal ligament stem cells in the inflammation group showed higher multiplication capacity, but the osteogenesis ability was lower compared with the normal control group. The expressions of Runx2 mRNA and Osterix mRNA were dropped significantly after the stimulus of tumor necrosis factor-α(P < 0.05), but the interleukin-1β and interleukin-6 did not have a significant impact. Tumor necrosis factor-α at 0.1 and 1 μg/L had no significant effects on the expression of Runx2 mRNA, but the expression of Runx2 mRNA was decreased significantly after treatment with 10 μg/L tumor necrosis factor-α (P < 0.05). It is confirmed that the molecular signaling mechanism inside the periodontal ligament stem cells is changed under inflammatory microenvironment, so that the differentiation capacity of cells from the inflammatory sources is lowered. Moreover, tumor necrosis factor-α is one of the key factors and its optimal concentration is 10 μg/L.
中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程 相似文献
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BACKGROUND: Articular cartilage injuries can result from a variety of causes. Conventional therapy cannot obtain the optimal clinical results. Low-intensity pulsed ultrasound has been shown to promote the repair of injured articular cartilage.
OBJECTIVE: To investigate the effects of low-intensity pulsed ultrasound on the repair of injured articular cartilage.
METHODS: Twenty New Zealand white rabbits were used to establish knee arthritis models and equally randomized into study and control groups, respectively. Rabbits in the study group received low-intensity pulsed ultrasound treatment, and sham low-intensity pulsed ultrasound treatment was given in the control group. At 8 weeks after treatment, pathological change and histological scores in articular cartilage tissue collected from both groups were determined. Moreover, the ultrastructure and type II collagen expression of chondrocytes were determined. Matrix metalloproteinase-13 mRNA expression was detected by quantitative real-time PCR.
RESULTS AND CONCLUSION: At 8 weeks after treatment, toluidine blue staining showed a disordered arrangement of cells, decreased number of cartilage cells in each layer and cluster in the control group. Light disordered arrangement of cells, decreased appearance of the superficial layer cells and the cluster phenomenon were observed in the study group. Articular cartilage tissue scores were significantly decreased in the study group compared with the control group (P < 0.05). The chondrocytes were small, enlarged intracellular mitochondria and rough endoplasmic reticulum, cytoplasmic swelling, collagen fibrils coarse, well developed Golgi apparatus, and nuclear fragmentation were observed in the control group. In addition, the normal structure of organelles disappeared and cell degeneration was observed in the control group. In the study group, the size of chondrocytes and the Golgi complex and other organelles were normal, and the protein polysaccharide granules were observed in the cytoplasm and membrane. The mRNA expression of matrix metalloproteinase-13 in the study group was significantly lower than that in the control group (P < 0.05). Type II collagen immunoreactivity in the study group was stronger than that in the control group. No incision infection, suppuration, red swelling appeared in all rabbits. Our results suggest that low-intensity pulsed ultrasound can be used for the treatment of articular cartilage injury by alleviating the degradation of collagen type II and inhibiting the expression of matrix metalloproteinase-13.
中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程 相似文献
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BACKGROUND: Previous studies have found that cryptotanshinone represses multiple tumors, but little is reported on its effect on renal carcinoma.
OBJECTIVE: To explore the effect of cryptotanshinone on the proliferation and apoptosis of the renal carcinoma stem cells.
METHODS: CD133+ renal carcinoma stem cells were separated from OS-RC-2 cells by immunomagnetic bead separation. Effects of 0, 0.2, 1, 5 mg/L cryptotanshinone on the proliferation and apoptosis of CD133+ renal carcinoma stem cells were detected by MTT and flow cytometry, respectively. Expression levels of Ki67, Bcl-2, Caspase-3 and p-Caspase-3 protein were detected by western blot assay.
RESULTS AND CONCLUSION: After magnetic cell sorting, the percentage of CD133+ cells was increased from 6.32% to 82.73%, and there was a significant difference before and after cell sorting (P < 0.001). Cryptotanshinone could repress the proliferation of CD133+ renal carcinoma stem cells and promote cell apoptosis in a dose-dependent manner. The protein expression levels of Ki67 and Bcl-2 in the 5 mg/L cryptotanshinone group were significantly decreased compared with the control group, while the protein expression level of p-Caspase-3 protein was significantly increased. In addition, there was no difference in the protein expression of Caspase-3 between cryptotanshinone and control group. These findings indicate that cryptotanshinone may be a potent anticancer drug for the treatment of renal carcinoma by inhibiting expression of Ki67 and Bcl-2 and promoting protein expression of p-Caspase-3.
中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程 相似文献