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BACKGROUND:Caffeic acid phenethyl ester (CAPE) can inhibit lipid peroxidation after rat brain injury. However, the trend of 5-lipoxygenaseis (5-LOX) and cysteinyl leukotrienes (CysLTs) in model of Parkinson’s disease, and whether CAPE protects against rotenone-induced cel ular injuries by inhibiting the levels of 5-LOX and CysLTs stil need further research. OBJECTIVE:To investigate the protective effect of CAPE on the rotenone-induced Parkinson-like injury, and to determine whether 5-LOX involved. METHODS:(1) PC12 cel s in good-growth were col ected and divided into five groups cultured with different concentrations of rotenone (0, 0.01, 0.1, 1, 10μmol/L). 24 and 48 hours later, changes of cel ular morphology and activity were observed to single out the optimum concentration of rotenone;at 24 hours, the levels of 5-LOX and CysLTs were detected by western blotting and ELISA, respectively. (2) PC12 cel s were pretreated with different concentrations of CAPE (0, 0.01, 0.1, 1, 10 μmol/L) for 30 minutes, and 1 μmol/L rotenone was then added. The other cel s received no intervention as blank control group. Subsequently, the cel activity was detected, and the CysLTs production was detected by ELISA at 24 hours. RESULTS AND CONCLUSION:(1) Rotenone (0.1-10μmol/L) could induce PC12 cel injury with overt morphological and cel activity changes at 24 hours, especial y the 1 μmol/L rotenone. (2) Rotenone also significantly increased the 5-LOX expression and CysLTs production in a concentration-dependant manner. (3) CAPE (1-10μmo/L) significantly attenuated rotenone-induced CysLTs production and cel viability reduction in a concentration-dependant manner. (4) These results suggest that CAPE protects against PC12 cel injuries in the model rat with Parkinson’s disease induced by rotenone involving 5-Lox.
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BACKGROUND:Brahma-related gene 1 (Brg1), a catalytic subunit of an important chromatin remodeling complex, has been considered as a key nuclear transcriptional factor, and tends to be decreased in diabetic cardiomyopathy. OBJECTIVE:To construct an adenovirus vector carrying Brg1, and observe its protective role in oxidative stress induced-cardiomyocyte apoptosis. METHODS:The recombinant adenovirus plasmid was linearized and transfected into HEK293 cel s using Fugene HD for packaging and amplification. The adenovirus particles were further purified, quantified, and sequential y transfected to cardiomyocytes of neonatal Sprague-Dawley rats. The Adeno-EGFP transfected and non-transfected cardiomyocytes were used as control group. 24 hours later, the transfection efficiency was observed by fluorescent microscope, and expressions of Brg1 mRNA and protein were detected by quantified PCR and western blotting. After treatment with 100 μmol/L H2O2 for 12 hours, the expressions of Brg1 protein and cleaved-Caspase 3 were measured by western blotting, and cel apoptosis was analyzed by flow cytometry. RESULTS AND CONCLUSION:(1) The recombinant adenovirus vector of Brg1 had been successful y transfected into cardiomyocytes with higher expressions of Brg1 mRNA and protein, and the transfection efficiency reached more than 90%. (2) After H2O2 treatment, the Brg1 was significantly down-regulated in contrast to the up-regulation of cleaved-Caspase 3;the flow cytometry data showed that the apoptotic cel s were increased. But in Adeno-Brg1 transfected cardiomyocytes, the H2O2 induced cel apoptosis was significantly decreased compared with non-transfected cel s and empty vector transfected cel s. (3) These results suggest that oxidative stress can directly inhibit the Brg1 expression, and overexpression of Brg1 can protect the cardiomyocytes from cel apoptosis induced by oxidative stress.
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Increasing studies have shown that endothelial progenitor cel s can not only promote the damaged endothelial cel s and vascular repair in ischemic brain tissue, but also promote neurogenesis, and thus promote neurological function recovery. Endothelial progenitor cel s transplantation may become an effective approach for the treatment of ischemic stroke.
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BACKGROUND:The linkage and synergistic effect of adaptor proteins can effectively regulate signal transduction of T cel s, which can form a limit or amplification cascade to realize the complex immune function of T cel s. C-terminal Src kinase (Csk)-binding protein (Cbp) is an adaptor protein, which mainly exert the negative feedback regulation of Src kinase activity. This negative feedback effect depends on Y317 of Cbp, which may be involved in the SH2 domain of Csk. OBJECTIVE:To explore the effects of high expression of Cbp on ultrastructure and related biological function of Jurkat cel s. METHODS:The virus particles were constructed with expressing enhanced green fluorescent protein (EGFP) only and Cbp-EGFP fusion protein to transfect Jurkat cel s. There were untransfected group (Jurkat group), negative control group (transfected with expression of EGFP virus only), and Cbp group (transfected with Cbp-EGFP virus). RESULTS AND CONCLUSION:Confocal microscope showed that cel transfection efficiency was more than 95%and Cbp was located on the cel membrane. Optical microscope showed after transfection with Cbp-EGFP virus, more Jurkat cel s shrunk, with poor size uniformity. Apoptosis detection showed that after transfection with Cbp-EGFP virus, the number of apoptotic and necrotic cel s was greatly increased. Cbp mRNA expression was increased, Csk expression was decreased obviously and lymphocyte-specific protein tyrosine kinase expression was increased. So, in Jurkat cel s, the high expression of Cbp can decrease the uniformity of cel s and increase the necrosis cel s, thus inhibiting the signal transduction.
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ObjectiveToinvestigatetheroleofimmuneinflammatoryreactionintheformationof intracranial aneurysm. Methods The intracranial aneurysms in 40 patients of craniotomy ( intracranial aneurysm group) and the vascular specimens in 20 craniotomy patients w ith traumatic brain injury (control group) w ere col ected. Fluorescence quantitative polymerase chain reaction w as used to detect the expression of interleukin (IL)-17 receptor in the arterial w al . Flow cytometry w as used to detect the Th-17 cel s in peripheral blood. Enzyme-linked immunosorbent assay w as used to measure the levels of IL-17, IL-6 in the arterial w al and tumor necrosis factor-α( TNF-α) in peripheral blood. Results There w ere no significant differences in the age (62.6 ±8.7 years vs.61.4 ±7.9 years;t=0.342;P=0.681), proportions of male (60.0%vs.65.0%; χ2 =0.246, P=0.434), hypertension ( 12.5%vs.10.0%; χ2 =0.315, P=0.492), diabetes (75.0%vs.10.0%; χ2 =0.284, P=0.482), and smoking (35.5%vs.30.0%; χ2 =0.224, P=0.413) betw een the intracranial aneurysms group and the control group. The expression of IL -17 receptor in the arterial w al (0.106 ±0.032 vs.0.264 ±0.071; t=5.115, P=0.001) and the proportion of Th17 cels in peripheral blood (2.75%±0.53%vs.7.18%±1.54%; t=8.436, P<0.001) and IL-17 level ( 7.32 ±1.82 μg/L vs.22.64 ±4.51 μg/L; t= 8.357, P< 0.001 ) in the control group w ere significantly low er than those in the intracranial aneurysm group. The levels of IL-6 (1.15 ±0.24 μg/L vs. 19.64 ±4.16 μg/L; t=9.527, P<0.001) and TNF-α(1.43 ±0.31 μg/L vs.26.17 ±4.32 μg/L; t=9.816, P<0.001) in the arterial wal in the control group were significantly lower than those in the intracranial aneurysm group. Conclusions The expression of IL-17 receptor in the arterial w al , the proportion of the Th17 cels and IL-17 level in peripheral blood were increased in patients with intracranial aneurysms. Immune inflammation may be involved in the formation of intracranial aneurysm.
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BACKGROUND:Hematopoietic microenvironment regulates hematopoietic stem cel s mainly through the cel-cel , cel-soluble regulator and cel-extracel uar matrix modes. OBJECTIVE:To review the molecular mechanism underlying regulation of hematopoietic stem cel s under the cel-cel , cel-soluble factors and cel-extracel ular matrix modes. METHODS:A computer-based online search of PubMed database was performed to search related articles with the key words of“Niche, HSC, VCAM-1, VLA-4, TPO, MPL, ECM, Integrin, N-cadherin, ANG-1, Tie2, VLA-5, Jagged-1, Notch, CXCL12, CXCR4, SCF, Kit, BMPs/TGF-β, TGF-βR, IFNα, IFNαR”in English. Irrelevant or repetitive studies as wel as old literature were excluded, and final y 80 articles were included in result analysis. RESULTS AND CONCLUSION:Cel s on the endosteal surface, vascular endothelial cel s, perivascular cel s, and some unknown or certain cel s or smal molecules in the bone marrow cavity constitute the specialized microenvironment for hematopoietic stem cel s. Hematopoietic stem cel s in the hematopoietic microenvironment remain a relatively steady state, which is the result of mutual contact of hematopoietic stem cel s and hematopoietic microenvironment under the regulation of some important molecules, such as Pten and osteopontin, via the pathway between cel-intercel ular adhesions, intercel ular soluble factors, and cel s and extracel ular matrix.
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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.
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BACKGROUND:Cdh1 has been shown to express in rat hippocampus and cortex in a large number. Moreover, in vitro test demonstrated that Cdh1 expression was higher in neurons than in neural stem cel s, which possibly associated with the differentiation of neural stem cel s into neurons. However, the effects of anaphase promoting complex Cdh1 on ischemic neuronal damage remain unclear. OBJECTIVE:To investigate the expression of Cdh1 and its downstream substrate in primary cultured neurons with oxygen-glucose deprivation. METHODS:Primary neurons from cortex of postnatal 24-hour rat pups were cultured in vitro, and identified by immunofluorescence staining. The oxygen-and glucose-deprived models were established by three gas incubator fil ed with nitrogen in sugar-free Earle’s solution. After 1 hour of hypoxia, reoxygenation was conducted. Real-time fluorescent quantitative PCR was used to detect the mRNA expression of Cdh1 and its downstream substrates Skp2, Cyclin B1 before hypoxia, 6 hours, 1, 3, 7 days after oxygen glucose deprivation. RESULTS AND CONCLUSION:After oxygen glucose deprivation, the expression of Cdh1 and Cyclin B1 in primary neurons was increased (P<0.05), while Skp2 expression was decreased (P<0.05). Above data indicated that Cdh1 expression in neurons increased after oxygen-glucose deprivation. It may degrade Skp2 and participate in hypoxic neuronal apoptosis by ubiquitination.
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BACKGROUND:Self-assembling peptide scaffolds have attracted much more attention among three dimensional biomaterials due to its biocompatibility, biodegradation, and tailor-made properties. OBJECTIVE:To review the structure and design of functionalized self-assembling RADA peptide, and the recent advances in the use of RADA self-assembling peptide for three-dimensional cel culture in cel therapy applications. METHODS:Total 224 literatures related to self-assembling peptide and tissue engineering from PubMed, Web of Science and CNKI databases were screened out for this review. The keywords were“self-assembly peptide, tissue engineering”in English and Chinese, respectively. Final y 48 of 224 articles about the design, fictionalization, and three-dimensional cel culture of peptide scaffolds were included in this review. RESULTS AND CONCLUSION:Self-assembling peptide could undergo spontaneous assembly into wel-ordered interwoven nanofibers in water and rapidly form hydrogel, which physical y mimics the architecture of natural extracel ular matrix to ensure a real three-dimensional microenvironment for cel s. In terms of bio-function, this material can be tailor-made with various bioactive short peptide motifs to promote cel adhesion, proliferation, and differentiation.
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BACKGROUND:Polyhydroxybutyrate-co-volerate (PHBV) is a noticeable tissue engineering material of polyhydroxyalkanoates family. It has the properties of low immune rejection response and good biocompatibility, and its degradation products are non-toxic. OBJECTIVE:To investigate the biocompatibility of PHBV membrane material and human bone marrow mesenchymal stem cel s in vitro. METHODS:Human bone marrow mesenchymal stem cel s at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cel number of two groups at 1, 2 and 4 hours and got the cel adherent rate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cel proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cel s at days 3, 6, 9 and 12 in both groups. After cel s were seeded upon PHBV membrane for 5 days, the cel growth upon the material was examined under a scanning electron microscope. RESULTS AND CONCLUSION:When the cel s were cultured for 1 hour, the adherent rate in the experimental group was lower than that in the control group;but there were no significant differences between two groups at the other two periods. No difference was found in the cel proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cel s seeded upon PHBV membrane for 5 days grew wel with spindle morphology and the intercel ular connections were tight and more extracel ular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cel s.
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BACKGROUND:Titanium artificial cervical disc has good biocompatibility, but titanium al oy is shown to have poor biological activity, low bonding strength, easy release of metal ions under physiological conditions. OBJECTIVE:To observe the effects of different coating for the titanium plate of domestic cervical artificial disc on adhesion and differentiation ability of rat bone marrow mesenchymal stem cel s. METHODS:Passage 3 bone marrow mesenchymal stem cel s from Wistar rats were seeded into 24-wel titanium plates with hydroxyapatite coating, titanium powder+hydroxyapatite coating and bare titanium plate. Cel culture was terminated after 24 and 48 hours, and the cel growth was observed under scanning electron microscope. After 24 hours of inoculation, osteogenic inducer was added;then, cel supernatant was col ected at 3, 5, 7 days after cel lysis and centrifugation to detect the activity of alkaline phosphatase. RESULTS AND CONCLUSION:After composite culture with titanium plates with hydroxyapatite coating or titanium powder+hydroxyapatite coating, cultured cel s were in polygonal shape, and pseudopodia were extended into the micropores that were adhered closely to the material surface. Cel s cultured with bare titanium plates had poor differentiation and adhesion rate. With time, the expression of alkaline phosphatase was increased in each group, especial y in the groups of titanium plates with hydroxyapatite coating and titanium powder+hydroxyapatite coating (P<0.05). These findings indicate that titanium plates with hydroxyapatite coating or titanium powder+hydroxyapatite coating can promote adhesion and differentiation of rat bone marrow mesenchymal stem cel s.
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BACKGROUND:Establishment of in vitro culture system of human placental microvascular endothelial cel s with high purity is very important. In recent studies, some scholars have successful y obtained a large number of placental microvascular endothelial cel s by three-stepenzyme digestion and magnetic separation method, but the procedures were extremely complex and it had great damage to the cel s. Therefore, how to separate human placental microvascular endothelial cel s easily and obtain high-purified cel s has become a research hotspot. OBJECTIVE:To investigate an efficient method to isolate and purify human placental microvascular endothelial cel s from early vil us microvessels, observe the cel growth and identify the cel s. METHODS:The vil i from normal early pregnancies (6-8 weeks) after artificial abortion were col ected aseptical y. Using two-step digestion procedure and discontinuous Percol density gradient centrifugation method, human placental microvascular endothelial cel s were obtained. Then the cel s were identified by trypsin digestion method and repeated adherence method. RESULTS AND CONCLUSION:Human placental microvascular endothelial cel s were isolated successful y from early vil i. The primary cel s adhered to the wal s after inoculated for 24 hours and entered logarithmic phase at 10 days. 80%of the cel s achieved a confluence at 12-13 days after inoculating. The subculture cel s grew swiftly with the typical cobblestone appearance. Immunofluorescence staining showed that, cultured human placental microvascular endothelial cel s demonstrated a strong positive reaction to von Wil ebrand factor antigen and CD31, accounting for 100%. MTT assay results showed that, human placental microvascular endothelial cel s at passage 5 exhibited an S-shaped growth curve. High-purity human placental microvascular endothelial cel s can be obtained by proteolytic enzymes digestion and discontinuous Percol density gradient centrifugation method, and the purity is detected by trypsin digestion method and repeated adherence method.
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BACKGROUND:Telomerase reverse transcriptase (TERT) plays an important role in telomerase activation, however there is rare report addressing the construction of the lentivirus targeted its genes to inhibit its expression in the spinal cord astrocytes. OBJECTIVE:To construct recombinant lentivirus vector expressing smal interfering RNA against TERT gene and to evaluate its potential for inhibiting the TERT expression. METHODS:After shRNA-TERT sequence was designed and synthesized, the sequence was amplified by PCR and then connected to plasmid pLentilox3.7U6-hTERT to construct recombinant plasmid. The recombinant plasmid was then transfected to DH5αcel s to screen positive colony, and the sequence was identified. The recombinant plasmid pLentilox3.7U6-TERT was transfected in 293T cel s, generating recombinant lentivirus Le-TERT. The titer of recombinant lentivirus was determined and Le-TERT was transfected into the rat spinal cord astrocytes. The expression of TERT in astrocytes was detected by RT-PCR, western blot and immunofluorescence assay. RESULTS AND CONCLUSION:The gene sequencing analysis confirmed that, recombinant plasmid pLentilox3.7U6-TERT was successful y constructed. The real-time quantitative PCR, western blot analysis and immunofluorescence assay indicated that, after Le-TERT was transfected in the astrocytes for 4 days, the inhibition rate of TERT mRNA was (63.98±2.6)%, and Le-TERT was lowly expressed in the transfected astrocytes. Recombinant expression vector pLentilox3.7U6-TERT can produce the lentivirus at high titer and effectively inhibit TERT expression in the transfected astrocytes.
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BACKGROUND:Clinical y, bone marrow mesenchymal stem cel s combined with artificial bones for early osteonecrosis of the femoral head have a wonderful outcome. OBJECTIVE:To review the biological properties of bone marrow mesenchymal stem cel s and to summarize the application progress of bone marrow mesenchymal stem cel s combined with different artificial bones in the treatment of early osteonecrosis of the femoral head. METHODS:PubMed (2003-2013), FMJS (2003-2013), Wanfang (2005-2013), CNKI (2005-2013) and CBM (2005-2013) databases were retrieved by computer using the keywords of“bone marrow mesenchymal stem cel s;artificial bone;osteonecrosis of the femoral head”in Chinese and English. RESULTS AND CONCLUSION:It wil lower the pressure of the femoral head, accelerate repair of the blood capil ary, improve the blood supply, osteoblast proliferation and differentiation, and thus delay or even prevent artificial joint replacement after osteonecrosis of the femoral head by applying bone marrow mesenchymal stem cel s combined with different artificial bones, such as corl ine hydroxyapatite, calcium hydroxylaptite, biological ceramics and calcium sulfate bones. But now, there are stil a lot of problems which need to be solved, including pathological mechanism underlying bone marrow mesenchymal stem cel s for treatment of osteonecrosis of the femoral head, obvious difference between the quantity and quality of seed cel s because of individual difference, different sites and culture techniques. So, artificial bone materials are under review, and large-sample randomized control trials are required. Its long-term outcomes also lack for fol ow-up observation, as wel as there is no a unified quantitative standard for the appropriate selection of indication, curative effect evaluation and the awareness of the operation.
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BACKGROUND:Kunming mouse embryonic fibroblasts are the most common feeder layers at present, and there are rare reports addressing C57BL/6 mouse embryonic fibroblasts as feeder layers. OBJECTIVE:To separate and culture C57BL/6 mouse embryonic fibroblasts in vitro, and produce feeder layers to enlarge the resources of mouse embryonic fibroblasts. METHODS:C57BL/6 mouse embryonic fibroblasts were isolated and cultured by trypsin digestion method in vitro. The biological characteristics and growth rule of the fibroblasts were investigated, then the feeder layers for the cel culture were produced. The growth of cel colonies on the prepared feeder layer was tested. RESULTS AND CONCLUSION:C57BL/6 mouse embryonic fibroblasts grew wel with a large amount, by trypsin digestion method at different concentrations. There was no significance in the survival rate after cryopreservation for 1 week, 2 weeks, 1 month, 3 months and 6 months. The cel s were proliferative from the second to fifth passage and declined sharply after the sixth passage. The planted mouse embryonic fibroblasts feeder layers had a high activity within 3 days, but got a sharp decline after 4 days. So it is best to use C57BL/6 mouse embryonic fibroblast feeder layers within 3 days after they’re inactivated. C57BL/6 mouse embryonic fibroblast feeder layer can support embryonic stem cel s and induce pluripotent stem cel s to grow as Kunming mouse embryonic fibroblasts.
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BACKGROUND:Bone marrow mesenchymal stem cel s have attracted widespread attention for the capabilities of self-renewal and muti-differentiation, which have been used in treatment of various diseases. OBJECTIVE:To study the effect of three-dimensional spheroid culture system on the stemness and senescence of bone marrow mesenchymal stem cel s. METHODS:Mesechyaml stem cel s were isolated from the bone marrow of C57/B6 mice, 3 weeks old, and cultured onto the culture plates coated with or without chitosan. After 5 days of culture, the cel phenotype and expression of stemness related markers CD44 and Sca-1 were analyzed by flow cytometry. PI and Annexin-V staining were used to detect cel apoptosis. Also,β-Gal staining was applied for identification of aging. RESULTS AND CONCLUSION:The mouse mesenchymal stem cel s began to form spheroids on day 3. The stemness-related markers, including CD44 and Sca-1, expressed higher in spheroid mesenchymal stem cel s than the cel s under normal culturing. Compared with the normal culture group, the apoptosis and senescence of cel s from spheroid culture were lower. The results indicate that the formation of spheroids on chitosan films can increase the stemness, decrease the apoptosis and slow the senescence of mesenchymal stem cel s.
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BACKGROUND:Previous studies have found that embryonic bone marrow mesenchymal stem cel s can promote human Th17 cel proliferation, but the inherent regulatory mechanisms stil need to be elucidated. OBJECTIVE:To investigate the role of Tol-like receptor 3 in the immunoregulation of Th17 cel s by mesenchymal stem cel s. METHODS:Human CD4+T cel s from healthy donors were isolated by immunomagnetic bead method, and then cultured alone or co-cultured with embryonic bone marrow mesenchymal stem cel s for 4 days. The mRNA expression level of interleukin-17, Tol-like receptor 3 and MyD88 was detected by real-time PCR. RESULTS AND CONCLUSION:Compared with CD4+T cel cultured alone group, the mRNA level of interleukin-17 was significantly higher in the co-culture group (3.59±0.11 vs. 1.14± 0.08, P<0.01). Consistent with the expression of interleukin-17 mRNA, increased level of Tol-like receptor 3 mRNA was detected in the co-culture group compared with the CD4+T cel cultured alone group (3.10±1.60 vs. 0.94± 0.01, P<0.05). Furthermore, MyD88 in the co-culture group was significantly higher than that in CD4+T cel cultured alone group (2.29±0.05 vs. 1.85±0.31, P<0.01). Tol-like receptor 3 may be involved in the immunoregulation of Th17 cel s by embryonic bone marrow mesenchymal stem cel s, which provides experimental evidence for potential cel therapeutic strategy.
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BACKGROUND:Studies have found that bone marrow mesenchymal stem cel s, under certain conditions, can be induced to differentiate into neurons and glial cel s, which to some extent solves the problem of the source of seed cel s. Induction methods currently used are different, and their efficiencies are not the same. OBJECTIVE:To observe the effects of different antioxidants on differentiation of rat bone marrow mesenchymal stem cel s into neuron-like cel s in vitro. METHODS:Bone marrow mesenchymal stem cel s from Wistar rats were divided into four groups:non-intervention group,β-mercaptoethanol group, retinoic acid group,β-mercaptoethanol+retinoic acid group. Changes in cel morphology and positive rate of neuron-specific enolase and microtubule-associated protein 2 were observed and detected at 5 hours, 12 hours, 1 day, 3 days, 5 days, 7 days, and 10 days after induction. RESULTS AND CONCLUSION:Except non-intervention group, bone marrow mesenchymal stem cel s in the other three groups were gradual y becoming spindle-shaped, and gave birth to many smal protrusions that were interconnected into a network, showing neuron-like cel morphology. Immunocytochemical staining showed that the efficiency of theβ-mercaptoethanol+retinoic acid group was the highest at 10 days after induction, and the positive rates of neuron-specific enolase and microtubule-associated protein 2 were 71.63%and 79.72%, respectively. The results show thatβ-mercaptoethanol can be combined with retinoic acid to accelerate the differentiation of bone marrow mesenchymal stem cel s into neuron-like cel s.
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BACKGROUND:Mesenchymal stem cel s can differentiate into nerve cel s by chemical induction or co-culture method, but whether mesenchymal stem cel s co-cultured with Schwann cel s differentiate into neuronal-like cel s or Schwann-like cel s is stil controversial. OBJECTIVE:To explore the inductive role of Schwann cel s derived from rats in the differentiation of human umbilical cord mesenchymal stem cel s. METHODS:Cocultures of human umbilical cord mesenchymal stem cel s (1×109/L) and Schwann cel s (1×109/L) from neonatal rats were performed using transwel culture dishes. After 2 weeks of cocultures, morphology of the cultured human umbilical cord mesenchymal stem cel s was observed, and the phenotypic changes of cel s were also detected with immumocytochemistry techniques. RESULTS AND CONCLUSION:After 2 weeks of cocultures, some differentiated cel s showed neuron-like morphology, and expressed nestin, NF-200 andβ-III-tubulin, but did not express Schwann cel special marker S100 and oligodendrocytes special marker MAB1580. These findings indicate that human umbilical cord mesenchymal stem cel s can transdifferentiate into neuronal-like cel s by cocultures with rat’s Schwann cel s.