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Xu T.-T.,Stem Cell Institute of Binzhou Medical College | Zhang H.-P.,Stem Cell Institute of Binzhou Medical College | Wang Y.-S.,Stem Cell Institute of Binzhou Medical College
Chinese Journal of Tissue Engineering Research | Year: 2013

BACKGROUND: Studies have shown that acidic fibroblast growth factor can be strongly expressed in the yolk sac hematopoietic cells and fetal liver hematopoietic cells and in the differentiation process of embryonic stem cells to hematopoietic stem cells. OBJECTIVE: To explore the effect of acidic fibroblast growth factor on hematopoietic differentiation of mice embryonic stem cells, to test and verify the regulation effect of acidic fibroblast growth factor in the formation of hematopoietic cells through adding the acidic fibroblast growth factor into the embryoid body culture stages. METHODS: The mouse embryonic stem cells were cultured and then digested into single cells using trypsin when they were in good conditions on the feeder layer. The embryoid bodies were prepared with hanging drop method and then continue suspension cultured for 3, 5, 7 and 9 days with 1, 2 and 5 μg/L acidic fibroblast growth factor. The expression of acidic fibroblast growth factor in embryonic stem cell and embryoid bodies was detected with immunofluorescence, and the Flk-1+ and CD133+ positive rates were detected with flow cytometry. RESULTS AND CONCLUSION: Acidic fibroblast growth factor showed positive expression in embryoid bodies. In the effect of 5 μg/L acidic fibroblast growth factor, Flk-1+ cells expression was increased with time increasing, the CD133+ cells expression patterns were similar to Flk-1+ cells. In the effect of 1 μg/L acidic fibroblast growth factor, Flk-1+ cells expression peaked at 7 days and then decreased, and the CD133+ cells showed similar expression patterns. But in the effect of 2 μg/L acidic fibroblast growth factor, Flk-1+ cells expression was increased at 5 days, then decreased, and increased again at 9 days; while CD133+ cells expression emerged a rising trend totality. Acidic fibroblast growth factor can promote the generation of Flk-1+ and CD133+ cells, suggesting that acidic fibroblast growth factor can effectively promote the amplification of embryonic body and the production and proliferation of hemangioblasts. Source


Zhang H.,Stem Cell Institute of Binzhou Medical College | Tan Y.,Stem Cell Institute of Binzhou Medical College | Xu T.,Stem Cell Institute of Binzhou Medical College | Wang Y.-S.,Stem Cell Institute of Binzhou Medical College
Chinese Journal of Clinical Oncology | Year: 2012

Objectives: To study the expression of the wnt and sonic hedgehog homolog (Shh) proteins in the endometrial carcinoma cell line HEC-1A; and to study the growth of HEC-1A cells treated with lithium chloride, an inhibitor of glycogen synthase kinase-3. Methods: The HEC-1A cells were cultivated in vitro. Immunofluorescence was used to determine the expression of wnt and Shh in the cells. 3-( 4, 5 - Dimethylthiazol - 2 - yl) -2, 5-diphenyltetrazolium bromide (MTT) was used to investigate the proliferation of HEC-1A cells treated with different concentrations of lithium chloride. In addition, real-time polymerase chain reaction (RT - PCR) was used to determine the expression level of the nuclear transcription factor Gli-1. Results: Wnt and Shh were expressed in the HEC-1A cells. The amount of Shh initially increased, and then decreased. Lithium chloride inhibited the growth of HEC-1A cells in a concentration-dependent manner. The RT-PCR results showed that the highest expression level of GH-1 was in the 24 h group, which corresponded to a change in the Shh level. Conclusion: The wnt signaling pathway can regulate the Shh expression. Moreover, lithium chloride can inhibit the proliferation of HEC-1A cells in a concentration-time- dependent manner. Source

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