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Objective To evaluate the biological effect of tumor necrosis factor-α(TNF-α) on the proliferation and multi-directional differentiation of stem cells from rat apical papilla(SCAP).Methods SCAP was extracted by combining enzyme digestion method with tissue block method.The cells were divided into control group(TNF-α 0 ng/mL) and experimental group(TNF-α 5,10,20,50 ng/mL).The ability of proliferation of SCAP was measured by MTT method.The ability of osteogenic/dentinogenic differentiation of SCAP was measured by alizarin red staining and quantitative real-time PCR.The ability of adipogenic of SCAP was measured by oil red O staining.The expression of vascular related genes of SCAP was measured by quantitative real-time PCR.Results SCAP was consistent with the characteristics of mesenchymal stem cells and possessed the ability of multi-directional differentiation.The MTT results showed that experimental group promoted the proliferation of SCAP in comparison with the control group.The difference was statistically significant(P<0.05),and 10 ng/mL was the optimum concentration.The results of alizarin red staining showed that with the increase of the concentration of TNF-α,the mineralized nodules in the experimental group gradually became smaller,and the number of the formation decreased gradually.The results of quantitative real-time PCR showed that the expression of OC,DMP-1 and DSPP in the experimental group was significantly lower than that of the control group at 3 and 7 days,in which the expression of OC was statistically significant different(P<0.05);at 14 days,the expression of OC,DMP-1 in the experimental group was significantly lower than that of the control group(P<0.05).The result of Oil red O staining showed that with the increase of the concentration of TNF-α,the lipid droplets formation in the experimental group gradually decreased.The result of quantitative real-time PCR showed that the expression of ANGPT1,VEGFA,PECAM-1 in the experimental group was significantly lower than that of the control group(P<0.05).Conclusion TNF-α might promote the proliferation and inhibit the multi-directional differentiation of SCAP.
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Objective To observe the promoting effect of hypoxia on proliferation of human bone marrow mesenchymal stem cells (BMMSCs) and maintaining their potential of multi-directional differentiation in vitro.Methods BMMSCs were isolated from bone marrow blood samples of patients with bone fracture, and cultured under hypoxic (group A) or normoxic (group B) conditions.The morphology, proliferation and osteogenic and adipogenic potential of the BMMSCs were observed.Results BMMSCs in the group A showed a long spindle shape and a fish shoal-like distribution, and were well-grown, while the morphology of cells in the group B appeared polygonal or flat.The quantity and growth rate of BMMSCs in the group A were increased compared with the group B (P< 0.05), with an osteogenic and adipogenic potential.Conclusions Hypoxia can promote the proliferation of BMMSCs in vitro and maintain their multi-directional differentiation potential.
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To explore the clinical ,pathological and immunohistochemical features of carcinoma with multi-directional differentiation derived from junction of bladder and prostate .We collected clinical data and tissue samples of three typical cases of carcinoma from our hospital .Routine preparation of slides and immunohistochem-ical methods combined with clinical data and pathological changes analysis were adopted .We found that all 3 ca-ses occurred in the elderly lesions involving the bladder and prostate .Dysuria was the main common symptom .All the cases had a history of chronic inflammation in urethral and prostate gland .The following up data showed that survival time in two of them were no more than 15 months.The third patient without chemotherapy who took drugs of bicalutamide,enantone and pamidronate still was alive after 28 months.The pathological changes of these cases had the common features with diversity .They all showed the structure with nests ,papillary,solid,adenoid,single or syncytial cells .The nuclear of cancer cell was enlarged with hyperchromatic feature .The mitotic figures were easily found.The metaplasia and atypical hyperplasia of prostate were all found .The immunohistochemistry results showed positive results for HCK,LCK,p53,p63,PSA,P504S and negative for vimentin and S -100.The average proliferation index of Ki-67 was 77%.
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Objective To observe the effect of the in vitro isolation, culture and identification of rabbit bone marrow-derived mesenchymal stem cells (BMSCs) and explore the differentiation potentials of BMSCs. Methods BMSCs were isolated and purified by density gradient centrifugation combined with attachment culture method. The biological characteristics of BMSCs were observed under an inverted microscope. The BMSCs were identified with HE staining and CD34/CD44 immunohistochemical staining. The biological activity of BMSCs was detected by osteogenic induction and steatogenic induction. Results BMSCs were isolated successfully in vitro and had good homogenicity. Immunohistochemical staining of CD34/CD44 showed strong positive expression of CD44 and negative expression of CD34. Fortis osteogenic ability and positive staining of ALP could be detected by osteogenic induction, and fortis steatogenic ability and positive staining of axungia could be detected by steatogenic induction. Conclusion BMSCs have the capability of multi-directional differentiation in vitro.
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@# As a multipotential stem cell, adipose-derived mesenchymal cell presents similar characters with bone marrow-derived menchymal stem cell in morphology and biology, and has the ability of directional differentiation to the all three germ layers. The adipose-derived mesenchymal stem cell could be harvested conveniently with slighter wound, and has a broad prospect in cell therapy and tissue engineering.