wangqian Pokročilý
Počet príspevkov : 115 Registration date : 28.11.2013
| Predmet: LMP1 activated the activity of cyclin D1 promoter by the EGFR and STAT3 pathway Po máj 19, 2014 9:02 am | |
| A re cent study has shown that neural progenitor cells isolated from developing human brain tissues are susceptible to CMV infection and ABT-888 分子量 undergo apoptosis following infection. However, the amount of neural cells obtainable from human brain tissues is limited. Pluripotent stem cells, including embryonic stem cells and induced pluripotent stem cells, are characterized by the ability to differentiate into tissues de rived from any of the three embryonic germ layers. Recent advances in the method to induce efficient differentiation of either ESCs or iPSCs into specific cell lineages offer an opportunity to establish model systems for viral infections of various cell types, including neural cells. Furthermore, differentiated cells derived from pluripotent stem cells are obtainable in potentially unlimited amounts.<br><br> Previous works revealed that while mouse ESCs are not susceptible to murine CMV, NSPCs that are differentiated from them are susceptible and their proliferation and differentiation Afatinib 構造 are suppressed by MCMV. Experi ments with human ESCs are, however, complicated with ethical problems. In this study, to analyze the pathological effects of HCMV on neural cells, we prepared NSPCs from human iPSCs and examined whether NSPCs are susceptible to HCMV infec tion. The results indicated that NSPCs are susceptible to HCMV infection and undergo apoptosis caused by mitochon drial dysfunction and endoplasmic reticulum stress. Methods Cells and viruses The human fetal lung fibroblast MRC5 was grown in Dulbeccos modified Eagles medium supplemented with 10% fetal bovine serum.<br><br> The human foreskin fibroblast cell line hTERT BJ1 immortalized with the human telomer ase reverse transcriptase was grown in a medium consisting of 4 parts of DMEM and 1 part of medium 199 AG-1478 溶解度 supplemented with 10% FBS, 1 mM sodium pyruvate, and 2 mM glutamine. HCMV laboratory strain Towne was propagated in hTERT BJ1 cells. The human iPSC line MRC iPS 25 that was established from MRC5 by retroviral vector mediated transduction of the c Myc, Oct 4, Klf4, and Sox2 genes were cultured on mitomycin C treated mouse embryonic fibroblasts in an iPSC medium consisting of Knockout DMEM F12 supplemented with non essential amino acids, glutamax I, 20% Knockout Serum Replacement, B mercaptoethanol and basic fibroblast growth fac tor.<br><br> Induced differentiation on iPSCs into neural stem cells MRC iPSC 25 cells cultured under feeder free condi tions were induced to differentiate into neural stem pro genitor cells by the method of dual inhibition of the SMAD signaling pathway described previously. In brief, feeder free iPSCs were treated with the mTeSR1 medium containing Y27632 and maintained with a daily medium change for 4 days. Then the medium was replaced with iPSC medium supplemented with SB431542 and Noggin. This date was designated day 0. On day 2, culture medium was replaced with a medium consisting of 3 parts of iPSC medium and 1 part of N2 medium supplemented with SB431542 and Noggin. On day 4, culture medium was re placed with a medium consisting of 1 part of iPSC medium and 1 part of N2 medium supplemented with SB431542 and Noggin. | |
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